Pyrrole inhibitors of S-nitrosoglutathione reductase as therapeutic agents

ABSTRACT

The present invention is directed to inhibitors of S-nitrosoglutathione reductase (GSNOR), pharmaceutical compositions comprising such GSNOR inhibitors, and methods of making and using the same.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.14/922,928, filed Oct. 26, 2015. U.S. application Ser. No. 14/922,928 isa continuation of U.S. application Ser. No. 14/598,062, filed Jan. 15,2015, now U.S. Pat. No. 9,180,119. U.S. application Ser. No. 14/598,062is a continuation of U.S. application Ser. No. 14/173,377, filed Feb. 5,2014, now U.S. Pat. No. 8,957,105. U.S. application Ser. No. 14/173,377is a continuation of U.S. application Ser. No. 13/057,220, filed Feb. 2,2011, now U.S. Pat. No. 8,673,961. U.S. application Ser. No. 13/057,220is a 35 U.S.C. §371 national phase application of InternationalApplication Serial No. PCT/US2009/053931, filed Aug. 14, 2009 (WO2010/019910). International Application Serial No. PCT/US2009/053931claims priority to U.S. Provisional Application Ser. No. 61/089,313,filed Aug. 15, 2008 and U.S. Provisional Application Ser. No.61/116,982, filed Nov. 21, 2008. Each of these applications isincorporated herein by reference in its entirety.

FIELD OF INVENTION

The present invention is directed to novel pyrrole inhibitors ofS-nitrosoglutathione reductase, pharmaceutical compositions comprisingsuch inhibitors, and methods of making and using the same.

BACKGROUND OF THE INVENTION

The chemical compound nitric oxide is a gas with chemical formula NO. NOis one of the few gaseous signaling molecules known in biologicalsystems, and plays an important role in controlling various biologicalevents. For example, the endothelium uses NO to signal surroundingsmooth muscle in the walls of arterioles to relax, resulting invasodilation and increased blood flow to hypoxic tissues. NO is alsoinvolved in regulating smooth muscle proliferation, platelet function,neurotransmission, and plays a role in host defense. Although nitricoxide is highly reactive and has a lifetime of a few seconds, it canboth diffuse freely across membranes and bind to many molecular targets.These attributes make NO an ideal signaling molecule capable ofcontrolling biological events between adjacent cells and within cells.

NO is a free radical gas, which makes it reactive and unstable, thus NOis short lived in vivo, having a half life of 3-5 seconds underphysiologic conditions. In the presence of oxygen, NO can combine withthiols to generate a biologically important class of stable NO adductscalled S-nitrosothiols (SNO's). This stable pool of NO has beenpostulated to act as a source of bioactive NO and as such appears to becritically important in health and disease, given the centrality of NOin cellular homeostasis (Stamler et al., Proc. Natl. Acad. Sci. USA,89:7674-7677 (1992)). Protein SNO's play broad roles in cardiovascular,respiratory, metabolic, gastrointestinal, immune and central nervoussystem function (Foster et al., 2003, Trends in Molecular MedicineVolume 9, Issue 4, April 2003, pages 160-168). One of the most studiedSNO's in biological systems is S-nitrosoglutathione (GSNO) (Gaston etal., Proc. Natl. Acad. Sci. USA 90:10957-10961 (1993)), an emerging keyregulator in NO signaling since it is an efficient trans-nitrosatingagent and appears to maintain an equilibrium with other S-nitrosatedproteins (Liu et al., 2001) within cells. Given this pivotal position inthe NO—SNO continuum, GSNO provides a therapeutically promising targetto consider when NO modulation is pharmacologically warranted.

In light of this understanding of GSNO as a key regulator of NOhomeostasis and cellular SNO levels, studies have focused on examiningendogenous production of GSNO and SNO proteins, which occurs downstreamfrom the production of the NO radical by the nitric oxide synthetase(NOS) enzymes. More recently there has been an increasing understandingof enzymatic catabolism of GSNO which has an important role in governingavailable concentrations of GSNO and consequently available NO andSNO's.

Central to this understanding of GSNO catabolism, researchers haverecently identified a highly conserved S-nitrosoglutathione reductase(GSNOR) (Jensen et al., Biochem J., 331:659-668 (1998); Liu et al.,Nature, 410:490-494 (2001)). GSNOR is also known asglutathione-dependent formaldehyde dehydrogenase (GS-FDH), alcoholdehydrogenase 3 (ADH-3) (Uotila and Koivusalo, Coenzymes and Cofactors,D. Dolphin, ed. pp. 517-551 (New York, John Wiley & Sons, 1989)), andalcohol dehydrogenase 5 (ADH-5). Importantly GSNOR shows greateractivity toward GSNO than other substrates (Jensen et al., 1998; Liu etal., 2001) and appears to mediate important protein and peptidedenitrosating activity in bacteria, plants, and animals. GSNOR appearsto be the major GSNO-metabolizing enzyme in eukaryotes (Liu et al.,2001). Thus, GSNO can accumulate in biological compartments where GSNORactivity is low or absent (e.g. airway lining fluid) (Gaston et al.,1993).

Yeast deficient in GSNOR accumulate S-nitrosylated proteins which arenot substrates of the enzyme, which is strongly suggestive that GSNOexists in equilibrium with SNO-proteins (Liu et al., 2001). Preciseenzymatic control over ambient levels of GSNO and thus SNO-proteinsraises the possibility that GSNO/GSNOR may play roles across a host ofphysiological and pathological functions including protection againstnitrosative stress wherein NO is produced in excess of physiologicneeds. Indeed, GSNO specifically has been implicated in physiologicprocesses ranging from the drive to breathe (Lipton et al., Nature,413:171-174 (2001)) to regulation of the cystic fibrosis transmembraneregulator (Zaman et al., Biochem Biophys Res Commun, 284:65-70 (2001),to regulation of vascular tone, thrombosis and platelet function (deBelder et al., Cardiovasc Res. 1994 May; 28(5):691-4. (1994); Z.Kaposzta, A et al., Circulation; 106(24): 3057-3062, 2002) as well ashost defense (de Jesus-Berrios et al., Curr. Biol., 13:1963-1968(2003)). Other studies have found that GSNOR protects yeast cellsagainst nitrosative stress both in vitro (Liu et al., 2001) and in vivo(de Jesus-Berrios et al., 2003).

Collectively data suggest GSNOR as a primary physiological ligand forthe enzyme S-nitrosoglutathione reductase (GSNOR), which catabolizesGSNO and consequently reduces available SNO's and NO in biologicalsystems (Liu et al., 2001), (Liu et al., Cell, (2004), 116(4), 617-628),and (Que et al., Science, 2005, 308, (5728):1618-1621). As such, thisenzyme plays a central role in regulating local and systemic bioactiveNO. Since perturbations in NO bioavailability has been linked to thepathogenesis of numerous disease states, including hypertension,atherosclerosis, thrombosis, asthma, gastrointestinal disorders,inflammation and cancer, agents that regulate GSNOR activity arecandidate therapeutic agents for treating diseases associated withnitric oxide imbalance.

Currently, there is a great need in the art for diagnostics,prophylaxis, ameliorations, and treatments for medical conditionsrelating to increased NO synthesis and/or increased NO bioactivity. Inaddition, there is a significant need for novel compounds, compositionsand methods for preventing, ameliorating, or reversing otherNO-associated disorders. The present invention satisfies these needs.

SUMMARY OF THE INVENTION

The present invention provides novel pyrrole compounds useful asS-nitrosoglutathione reductase (“GSNOR”) inhibitors. The inventionencompasses pharmaceutically acceptable salts, prodrugs, and metabolitesof the described GSNOR inhibitors. Also encompassed by the invention arepharmaceutical compositions comprising at least one GSNOR inhibitor andat least one pharmaceutically acceptable carrier.

The compositions of the present invention can be prepared in anysuitable pharmaceutically acceptable dosage form.

The present invention provides a method for inhibitingS-nitrosoglutathione reductase in a subject in need thereof. Such amethod comprises administering a therapeutically effective amount of apharmaceutical composition comprising at least one GSNOR inhibitor or apharmaceutically acceptable salt thereof, a prodrug or metabolitethereof, in combination with at least one pharmaceutically acceptablecarrier. The GSNOR inhibitor can be a novel compound according to theinvention, or it can be a known compound which previously was not knownto be an inhibitor of GSNOR.

The present invention also provides a method of treating a disorderameliorated by NO donor therapy in a subject in need thereof. Such amethod comprises administering a therapeutically effective amount of apharmaceutical composition comprising at least one GSNOR inhibitor or apharmaceutically acceptable salt thereof, a prodrug, or metabolitethereof, in combination with at least one pharmaceutically acceptablecarrier. The GSNOR inhibitor can be a novel compound according to theinvention, or it can be a known compound which previously was not knownto be an inhibitor of GSNOR.

The present invention also provides a method of treating a cellproliferative disorder in a subject in need thereof. Such a methodcomprises administering a therapeutically effective amount of apharmaceutical composition comprising at least one GSNOR inhibitor or apharmaceutically acceptable salt thereof, a prodrug, or metabolitethereof, in combination with at least one pharmaceutically acceptablecarrier. The GSNOR inhibitor can be a novel compound according to theinvention, or it can be a known compound which previously was not knownto be an inhibitor of GSNOR.

The methods of the invention encompass administration with one or moresecondary active agents. Such administration can be sequential or in acombination composition.

Although methods and materials similar or equivalent to those describedherein can be used in the practice or testing of the present invention,suitable methods and materials are described below. All publiclyavailable publications, patent applications, patents, and otherreferences mentioned herein are incorporated by reference in theirentirety. In the case of conflict, the present specification, includingdefinitions, will control.

Both the foregoing summary and the following detailed description areexemplary and explanatory and are intended to provide further details ofthe compositions and methods as claimed. Other objects, advantages, andnovel features will be readily apparent to those skilled in the art fromthe following detailed description.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS A. Overview of theInvention

Until recently, S-nitrosoglutathione reductase (GSNOR) was known tooxidize the formaldehyde glutathione adduct, S-hydroxymethylglutathione.GSNOR has since been identified in a variety of bacteria, yeasts, plantsand animals and is well conserved. The proteins from E. coli, S.cerevisiae and mouse macrophages share over 60% amino acid sequenceidentity. GSNOR activity (i.e., decomposition of S-nitrosoglutathionewhen NADH is present as a required cofactor) has been detected in E.coli, in mouse macrophages, in mouse endothelial cells, in mouse smoothmuscle cells, in yeasts, and in human HeLa, epithelial and monocytecells. Human GSNOR nucleotide and amino acid sequence information can beobtained from the National Center for Biotechnology Information (NCBI)databases under Accession Nos. M29872, NM_000671. Mouse GSNOR nucleotideand amino acid sequence information can be obtained from NCBI databasesunder Accession Nos. NM_007410. In the nucleotide sequence, the startsite and stop site are underlined. CDS designates coding sequence. SNPdesignates single nucleotide polymorphism. Other related GSNORnucleotide and amino acid sequences, including those of other species,can be found in U.S. Patent Application 2005/0014697.

In accord with the present invention, GSNOR has been shown to functionin vivo and in vitro to metabolize S-nitrosoglutathione (GSNO) andprotein S-nitrosothiols (SNOs) to modulate NO bioactivity, bycontrolling the intracellular levels of low mass NO donor compounds andpreventing protein nitrosylation from reaching toxic levels.

Based on this, it follows that inhibition of this enzyme potentiatesbioactivity in all diseases in which NO donor therapy is indicated,inhibits the proliferation of pathologically proliferating cells, andincreases NO bioactivity in diseases where this is beneficial.

The present invention provides pharmaceutical agents that are potentinhibitors of GSNOR. In particular, provided are substituted pyrroleanalogs that are inhibitors of GSNOR having the structures depictedbelow (Formulas I and II), or a pharmaceutically acceptable salt,stereoisomer, or prodrug thereof.

Tri-substituted pyrrole analogs are potent inhibitors of GSNOR. As usedin this context, the term “analog” refers to a compound having similarchemical structure or function as compounds of Formula I-II that retainsthe pyrrole ring.

Some pyrrole analogs of the invention can also exist in various isomericforms, including configurational, geometric and conformational isomers,as well as existing in various tautomeric forms, particularly those thatdiffer in the point of attachment of a hydrogen atom. As used herein,the term “isomer” is intended to encompass all isomeric forms of acompound including tautomeric forms of the compound.

Illustrative compounds having asymmetric centers can exist in differentenantiomeric and diastereomeric forms. A compound can exist in the formof an optical isomer or a diastereomer. Accordingly, the inventionencompasses compounds in the forms of their optical isomers,diastereomers and mixtures thereof, including racemic mixtures.

It should be noted that if there is a discrepancy between a depictedstructure and a name given to that structure, the depicted structurecontrols. In addition, if the stereochemistry of a structure or aportion of a structure is not indicated with, for example, bold, wedged,or dashed lines, the structure or portion of the structure is to beinterpreted as encompassing all stereoisomers of the described compound.

In accordance with the invention, the levels of the S-nitrosoglutathionereductase in the biological sample can be determined by the methodsdescribed in U.S. Patent Application Publication No. 2005/0014697. Theterm “biological sample” includes, but is not limited to, samples ofblood (e.g., serum, plasma, or whole blood), urine, saliva, sweat,breast milk, vaginal secretions, semen, hair follicles, skin, teeth,bones, nails, or other secretions, body fluids, tissues, or cells.

B. Definitions

As used herein, “about” will be understood by persons of ordinary skillin the art and will vary to some extent on the context in which it isused. If there are uses of the term which are not clear to persons ofordinary skill in the art given the context in which it is used, “about”will mean up to plus or minus 10% of the particular term.

The term “acyl” includes compounds and moieties that contain the acetylradical (CH₃CO—) or a carbonyl group to which a straight or branchedchain lower alkyl residue is attached.

The term “alkyl” as used herein refers to a straight or branched chain,saturated hydrocarbon having the indicated number of carbon atoms. Forexample, (C₁-C₆) alkyl is meant to include, but is not limited tomethyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, pentyl,isopentyl, neopentyl, hexyl, isohexyl, and neohexyl. An alkyl group canbe unsubstituted or optionally substituted with one or more substituentsas described herein.

The term “alkenyl” as used herein refers to a straight or branched chainunsaturated hydrocarbon having the indicated number of carbon atoms andat least one double bond. Examples of a (C₂-C₈) alkenyl group include,but are not limited to, ethylene, propylene, 1-butylene, 2-butylene,isobutylene, sec-butylene, 1-pentene, 2-pentene, isopentene, 1-hexene,2-hexene, 3-hexene, isohexene, 1-heptene, 2-heptene, 3-heptene,isoheptene, 1-octene, 2-octene, 3-octene, 4-octene, and isooctene. Analkenyl group can be unsubstituted or optionally substituted with one ormore substituents as described herein.

The term “alkynyl” as used herein refers to a straight or branched chainunsaturated hydrocarbon having the indicated number of carbon atoms andat least one triple bond. Examples of a (C₂-C₈) alkynyl group include,but are not limited to, acetylene, propyne, 1-butyne, 2-butyne,1-pentyne, 2-pentyne, 1-hexyne, 2-hexyne, 3-hexyne, 1-heptyne,2-heptyne, 3-heptyne, 1-octyne, 2-octyne, 3-octyne and 4-octyne. Analkynyl group can be unsubstituted or optionally substituted with one ormore substituents as described herein.

The term “alkoxy” as used herein refers to an —O-alkyl group having theindicated number of carbon atoms. For example, a (C₁-C₆) alkoxy groupincludes —O-methyl, —O-ethyl, —O-propyl, —O-isopropyl, —O-butyl,—O-sec-butyl, —O-tert-butyl, —O-pentyl, —O-isopentyl, —O-neopentyl,—O-hexyl, —O-isohexyl, and —O-neohexyl.

The term “aminoalkyl” as used herein, refers to an alkyl group(typically one to six carbon atoms) wherein one or more of the C₁-C₆alkyl group's hydrogen atoms is replaced with an amine of formula—N(R^(c))₂, wherein each occurrence of R^(c) is independently —H or(C₁-C₆) alkyl. Examples of aminoalkyl groups include, but are notlimited to, —CH₂NH₂, —CH₂CH₂NH₂—, —CH₂CH₂CH₂NH₂, —CH₂CH₂CH₂CH₂NH₂,—CH₂CH₂CH₂CH₂CH₂NH₂, —CH₂CH₂CH₂CH₂CH₂CH₂NH₂, —CH₂CH₂CH₂N(CH₃)₂,t-butylaminomethyl, isopropylaminomethyl and the like.

The term “aryl” as used herein refers to a 5- to 14-membered monocyclic,bicyclic or tricyclic aromatic ring system. Examples of an aryl groupinclude phenyl and naphthyl. An aryl group can be unsubstituted oroptionally substituted with one or more substituents as described hereinbelow. Examples of aryl groups include phenyl or aryl heterocycles suchas, pyrrole, furan, thiophene, thiazole, isothiazole, imidazole,triazole, tetrazole, pyrazole, oxazole, isoxazole, pyridine, pyrazine,pyridazine, and pyrimidine, and the like.

As used herein, the term “bioactivity” indicates an effect on one ormore cellular or extracellular process (e.g., via binding, signaling,etc.) which can impact physiological or pathophysiological processes.

The term “carbonyl” or “carboxy” or “carboxyl” includes compounds andmoieties which contain a carbon connected with a double bond to anoxygen atom. Examples of moieties containing a carbonyl include, but arenot limited to, aldehydes, ketones, carboxylic acids, amides, esters,anhydrides, etc.

The term “C_(m)-C_(n)” means “m” number of carbon atoms to “n” number ofcarbon atoms. For example, the term “C₁-C₆” means one to six carbonatoms (C₁, C₂, C₃, C₄, C₅ or C₆). The term “C₂-C₆” includes two to sixcarbon atoms (C₂, C₃, C₄, C₅ or C₆). The term “C₃-C₆” includes three tosix carbon atoms (C₃, C₄, C₅ or C₆).

The term “cycloalkyl” as used herein refers to a 3- to 14-memberedsaturated or unsaturated non-aromatic monocyclic, bicyclic or tricyclichydrocarbon ring system. Included in this class are cycloalkyl groupswhich are fused to a benzene ring. Representative cycloalkyl groupsinclude, but are not limited to, cyclopropyl, cyclobutyl, cyclobutenyl,cyclopentyl, cyclopentenyl, cyclopentadienyl, cyclohexyl, cyclohexenyl,1,3-cyclohexadienyl, cycloheptyl, cycloheptenyl, 1,3-cycloheptadienyl,1,4-cycloheptadienyl, -1,3,5-cycloheptatrienyl, cyclooctyl,cyclooctenyl, 1,3-cyclooctadienyl, 1,4-cyclooctadienyl,-1,3,5-cyclooctatrienyl, decahydronaphthalene, octahydronaphthalene,hexahydronaphthalene, octahydroindene, hexahydroindene, tetrahydroinden,decahydrobenzocycloheptene, octahydrobenzocycloheptene,hexahydrobenzocycloheptene, tetrahydrobenzocyclopheptene,dodecahydroheptalene, decahydroheptalene, octahydroheptalene,hexahydroheptalene, and tetrahydroheptalene,(1s,3s)-bicyclo[1.1.0]butane, bicyclo[1.1.1]pentane,bicyclo[2.1.1]hexane, Bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane,bicyclo[3.1.1]heptane, bicyclo[3.2.1]octane, bicyclo[3.3.1]nonane,bicyclo[3.3.2]decane, bicyclo[3.3.]undecane, bicyclo[4.2.2]decane,bicyclo[4.3.1]decane. A cycloalkyl group can be unsubstituted oroptionally substituted with one or more substituents as described hereinbelow.

The term “halogen” includes fluorine, bromine, chlorine, iodine, etc.

The term “haloalkyl” as used herein, refers to a C₁-C₆ alkyl groupwherein from one or more of the C₁-C₆ alkyl group's hydrogen atom isreplaced with a halogen atom, which can be the same or different.Examples of haloalkyl groups include, but are not limited to,trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl,pentachloroethyl, and 1,1,1-trifluoro-2-bromo-2-chloroethyl.

The term “heteroalkyl” by itself or in combination with another term,means, unless otherwise stated, a stable straight or branched chainalkyl, or combinations thereof, consisting of carbon atoms and from oneto three heteroatoms selected from the group consisting of O, N and S,and wherein the nitrogen and sulfur atoms may optionally be oxidized andthe nitrogen heteroatom may optionally be quaternized. The heteroatom(s)O, N and S can be placed at any position of the heteroalkyl group.Examples include —CH₂—CH₂—O—CH₃, —CH₂—CH₂—NH—CH₃, —CH₂—CH₂—N(CH₃)—CH₃,—CH₂—S—CH₂—CH₃, —CH₂—CH₂—S(O)—CH₃, —CH₂—CH₂—S(O)₂—CH₃, and—CH₂—CH═N—OCH₃. Up to two heteroatoms can be consecutive, such as, forexample, —CH₂—NH—OCH₃. When a prefix such as (C₂-C₈) is used to refer toa heteroalkyl group, the number of carbons (2 to 8, in this example) ismeant to include the heteroatoms as well. For example, a C₂-heteroalkylgroup is meant to include, for example, —CH₂OH (one carbon atom and oneheteroatom replacing a carbon atom) and —CH₂SH.

To further illustrate the definition of a heteroalkyl group, where theheteroatom is oxygen, a heteroalkyl group can be an oxyalkyl group. Forinstance, (C₂-C₅) oxyalkyl is meant to include, for example —CH₂—O—CH₃(a C₃-oxyalkyl group with two carbon atoms and one oxygen replacing acarbon atom), —CH₂CH₂CH₂CH₂OH, —OCH₂CH₂OCH₂CH₂OH, —OCH₂CH(OH)CH₂OH, andthe like.

The term “heteroaryl” as used herein refers to an aromatic heterocyclering of 5 to 14 members and having at least one heteroatom selected fromnitrogen, oxygen and sulfur, and containing at least 1 carbon atom,including monocyclic, bicyclic, and tricyclic ring systems.Representative heteroaryls are triazolyl, tetrazolyl, oxadiazolyl,pyridyl, furyl, benzofuranyl, thienyl (thiophen-yl), benzothienyl,quinolinyl, pyrrolyl, indolyl, oxazolyl, benzoxazolyl, imidazolyl,benzimidazolyl, thiazolyl, benzothiazolyl, isoxazolyl, pyrazolyl,isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl,cinnolinyl, phthalazinyl, quinazolinyl, pyrimidyl, azepinyl, oxepinyl,quinoxalinyl and oxazolyl. A heteroaryl group can be unsubstituted oroptionally substituted with one or more substituents as described hereinbelow.

As used herein, the term “heteroatom” is meant to include oxygen (O),nitrogen (N), and sulfur (S).

As used herein, the term “heterocycle” refers to 3- to 14-membered ringsystems which are either saturated, unsaturated, or aromatic, and whichcontains from 1 to 4 heteroatoms independently selected from nitrogen,oxygen and sulfur, and wherein the nitrogen and sulfur heteroatoms canbe optionally oxidized, and the nitrogen heteroatom can be optionallyquaternized, including, including monocyclic, bicyclic, and tricyclicring systems. The bicyclic and tricyclic ring systems may encompass aheterocycle or heteroaryl fused to a benzene ring. The heterocycle canbe attached via any heteroatom or carbon atom, where chemicallyacceptable. Heterocycles include heteroaryls as defined above.Representative examples of heterocycles include, but are not limited to,aziridinyl, oxiranyl, thiiranyl, triazolyl, tetrazolyl, azirinyl,diaziridinyl, diazirinyl, oxaziridinyl, azetidinyl, azetidinonyl,oxetanyl, thietanyl, piperidinyl, piperazinyl, morpholinyl, pyrrolyl,oxazinyl, thiazinyl, diazinyl, dioxanyl, triazinyl, tetrazinyl,imidazolyl, tetrazolyl, pyrrolidinyl, isoxazolyl, furanyl, furazanyl,pyridinyl, oxazolyl, benzoxazolyl, benzisoxazolyl, thiazolyl,benzthiazolyl, thienyl, pyrazolyl, triazolyl, pyrimidinyl,benzimidazolyl, isoindolyl, indazolyl, benzodiazolyl, benzotriazolyl,benzoxazolyl, benzisoxazolyl, purinyl, indolyl, isoquinolinyl,quinolinyl and quinazolinyl. A heterocycle group can be unsubstituted oroptionally substituted with one or more substituents as described hereinbelow.

The term “heterocycloalkyl” by itself or in combination with otherterms, represents, unless otherwise stated, cyclic versions of“heteroalkyl.” Additionally, a heteroatom can occupy the position atwhich the heterocycle is attached to the remainder of the molecule.Examples of heterocycloalkyl include 1-(1,2,5,6-tetrahydropyridyl),1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl,3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl,tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl,2-piperazinyl, and the like.

The term “hydroxyalkyl” as used herein, refers to an alkyl group havingthe indicated number of carbon atoms wherein one or more of the hydrogenatoms in the alkyl group is replaced with an —OH group. Examples ofhydroxyalkyl groups include, but are not limited to, —CH₂OH, —CH₂CH₂OH,—CH₂CH₂CH₂OH, —CH₂CH₂CH₂CH₂OH, —CH₂CH₂CH₂CH₂CH₂OH,—CH₂CH₂CH₂CH₂CH₂CH₂OH, and branched versions thereof.

The term “hydroxy” or “hydroxyl” includes groups with an —OH or —O⁻.

As used herein and unless otherwise indicated, the term “stereoisomer”means one stereoisomer of a compound that is substantially free of otherstereoisomers of that compound. For example, a stereomerically purecompound having one chiral center will be substantially free of theopposite enantiomer of the compound. A stereomerically pure compoundhaving two chiral centers will be substantially free of otherdiastereomers of the compound. In some embodiments, a stereomericallypure compound comprises greater than about 80% by weight of onestereoisomer of the compound and less than about 20% by weight of otherstereoisomers of the compound, for example greater than about 90% byweight of one stereoisomer of the compound and less than about 10% byweight of the other stereoisomers of the compound, or greater than about95% by weight of one stereoisomer of the compound and less than about 5%by weight of the other stereoisomers of the compound, or greater thanabout 97% by weight of one stereoisomer of the compound and less thanabout 3% by weight of the other stereoisomers of the compound.

As used herein, “protein” is used synonymously with “peptide,”“polypeptide,” or “peptide fragment.” A “purified” polypeptide, protein,peptide, or peptide fragment is substantially free of cellular materialor other contaminating proteins from the cell, tissue, or cell-freesource from which the amino acid sequence is obtained, or substantiallyfree from chemical precursors or other chemicals when chemicallysynthesized.

As used herein, “modulate” is meant to refer to an increase or decreasethe levels of a peptide or a polypeptide, or to increase or decrease thestability or activity of a peptide or a polypeptide. The term “inhibit”is meant to refer to a decrease in the levels of a peptide or apolypeptide or to decrease in the stability or activity of a peptide ora polypeptide. In preferred embodiments, the peptide which is modulatedor inhibited is S-nitrosoglutathione (GSNO) or protein S-nitrosothiols(SNOs).

As used here, the terms “nitric oxide” and “NO” encompass unchargednitric oxide and charged nitric oxide species, particularly includingnitrosonium ion (NO⁺) and nitroxyl ion (NO⁻). The reactive form ofnitric oxide can be provided by gaseous nitric oxide. Compounds havingthe structure X—NO_(y) wherein X is a nitric oxide releasing, deliveringor transferring moiety, including any and all such compounds whichprovide nitric oxide to its intended site of action in a form active fortheir intended purpose, and Y is 1 or 2.

As utilized herein, the term “pharmaceutically acceptable” meansapproved by a regulatory agency of a federal or a state government orlisted in the U.S. Pharmacopoeia or other generally recognizedpharmacopoeia for use in animals and, more particularly, in humans. Theterm “carrier” refers to a diluent, adjuvant, excipient, or vehicle withwhich the therapeutic is administered and includes, but is not limitedto such sterile liquids as water and oils.

A “pharmaceutically acceptable salt” or “salt” of a GSNOR inhibitor is aproduct of the disclosed compound that contains an ionic bond, and istypically produced by reacting the disclosed compound with either anacid or a base, suitable for administering to a subject. Apharmaceutically acceptable salt can include, but is not limited to,acid addition salts including hydrochlorides, hydrobromides, phosphates,sulphates, hydrogen sulphates, alkylsulphonates, arylsulphonates,arylalkylsulfonates, acetates, benzoates, citrates, maleates, fumarates,succinates, lactates, and tartrates; alkali metal cations such as Li,Na, K, alkali earth metal salts such as Mg or Ca, or organic aminesalts.

A “pharmaceutical composition” is a formulation comprising the disclosedcompounds in a form suitable for administration to a subject. Apharmaceutical composition of the invention is preferably formulated tobe compatible with its intended route of administration. Examples ofroutes of administration include, but are not limited to, oral andparenteral, e.g., intravenous, intradermal, subcutaneous, inhalation,topical, transdermal, transmucosal, and rectal administration.

The term “substituted” as used herein, means that any one or morehydrogens on the designated atom is replaced with a selection from theindicated group, provided that the designated atom's normal valency isnot exceeded, and that the substitution results in a stable compound.When a substituent is keto (i.e., ═O), then 2 hydrogens on the atom arereplaced. Ring double bonds, as used herein, are double bonds that areformed between two adjacent ring atoms (e.g., C═C, C═N, or N═N).

Substituents for the groups referred to as alkyl, heteroalkyl, alkylene,alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl andheterocycloalkenyl can be selected from a variety of groups including—OR^(d)′, ═O, ═NR^(d)′, ═N—OR^(d)′, —NR^(d)′R^(d)″, —SR^(d)′, -halo,—SiR^(d)′R^(d)″R^(d)′″, —OC(O)R^(d)′, —C(O)R^(d)′, —CO₂R^(d)′,—CONR^(d)′R^(d)″, —OC(O)NR^(d)′R^(d)″, —NR^(d)″C(O)R^(d)′,—NR^(d)′″C(O)NR^(d)′R^(d)″, —NR^(d)′″SO₂NR^(d)′R^(d)″,—NR^(d)″CO₂R^(d)′, —NHC(NH₂)═NH, —NR^(d)′C(NH₂)═NH, —NHC(NH₂)═NR^(d)′,—S(O)R^(d)′, —SO₂R^(d)′, —SO₂NR^(d)′R^(d)″, —NR^(d)″SO₂R^(d)′, —CN and—NO₂, in a number ranging from zero to three, with those groups havingzero, one or two substituents being exemplary.

R^(d)′, R^(d)″ and R^(d)′″ each independently refer to hydrogen,unsubstituted (C₁-C₈)alkyl, unsubstituted hetero(C₁-C₈) alkyl,unsubstituted aryl and aryl substituted with one to three substituentsselected from -halo, unsubstituted alkyl, unsubstituted alkoxy,unsubstituted thioalkoxy and unsubstituted aryl (C₁-C₄)alkyl. WhenR^(d)′ and R^(d)″ are attached to the same nitrogen atom, they can becombined with the nitrogen atom to form a 5-, 6- or 7-membered ring. Forexample, —NR^(d)′R^(d)″ can represent 1-pyrrolidinyl or 4-morpholinyl.

Typically, an alkyl or heteroalkyl group will have from zero to threesubstituents, with those groups having two or fewer substituents beingexemplary of the present invention. An alkyl or heteroalkyl radical canbe unsubstituted or monosubstituted. In some embodiments, an alkyl orheteroalkyl radical will be unsubstituted.

Exemplary substituents for the alkyl and heteroalkyl radicals includebut are not limited to —OR^(d)′, ═O, ═NR^(d)′, ═N—OR^(d)′,—NR^(d)′R^(d)″, —SR^(d)′, -halo, —SiR^(d)′R^(d)″R^(d)′″, —OC(O)R^(d)′,—C(O)R^(d)′, —CO₂R^(d)′, —CONR^(d)′R^(d)″, —OC(O)NR^(d)′R^(d)″,—NR^(d)″C(O)R^(d)′, —NR^(d)′″C(O)NR^(d)′R^(d)″,—NR^(d)′″SO₂NR^(d)′R^(d)″, —NR^(d)″CO₂R^(d)′, —NHC(NH₂)═NH,—NR^(a)′C(NH₂)═NH, —NHC(NH₂)═NR^(d)′, —S(O)R^(d)′, —SO₂R^(d)′,—SO₂NR^(d)′R^(d)″, —NR^(d)″SO₂R^(d)′, —CN and —NO₂, where R^(d)′, R^(d)″and R^(d)′″ are as defined above. Typical substituents can be selectedfrom: —OR^(d)′, ═O, —NR^(d)′R^(d)″, -halo, —OC(O)R^(d)′, —CO₂R^(d)′,—C(O)NR^(d)′R^(d)″, —OC(O)NR^(d)′R^(d)″, —NR^(d)″C(O)R^(d)′,—NR^(d)″CO₂R^(d)′, —NR^(d)′″SO₂NR^(d)′R^(d)″, —SO₂R^(d)′,—SO₂NR^(d)′R^(d)″, —NR^(d)″SO₂R^(d)′, —CN and —NO₂.

Similarly, substituents for the aryl and heteroaryl groups are variedand selected from: -halo, —OR^(e)′, —OC(O)R^(e)′, —NR^(e)′R^(e)″,—SR^(e)′, —R^(e)′, —CN, —NO₂, —CO₂R^(e)′, —C(O)NR^(e)′R^(e)″,—C(O)R^(e)′, —OC(O)NR^(e)′R^(e)″, —NR^(e)″C(O)R^(e)′, —NR^(e)″CO₂R^(e)′,—NR^(e)′″C(O)NR^(e)′R^(e)″, —NR^(e)′″SO₂NR^(e)′R^(e)″, —NHC(NH₂)═NH,—NR^(e)′C(NH₂)═NH, —NH—C(NH₂)═NR^(e)′, —S(O)R^(e)′, —SO₂R^(e)′,—SO₂NR^(e)′R^(e)″, —NR^(e)″SO₂R^(e)′, —N₃, —CH(Ph)₂, perfluoroalkoxy andperfluoro(C₁-C₄)alkyl, in a number ranging from zero to the total numberof open valences on the aromatic ring system.

R^(e)′, R^(e)″ and R^(e)′″ are independently selected from hydrogen,unsubstituted (C₁-C₈) alkyl, unsubstituted hetero(C₁-C₈) alkyl,unsubstituted aryl, unsubstituted heteroaryl, unsubstituted aryl(C₁-C₄)alkyl and unsubstituted aryloxy(C₁-C₄) alkyl. Typically, an aryl orheteroaryl group will have from zero to three substituents, with thosegroups having two or fewer substituents being exemplary in the presentinvention. In one embodiment of the invention, an aryl or heteroarylgroup will be unsubstituted or monosubstituted. In another embodiment,an aryl or heteroaryl group will be unsubstituted.

Two of the substituents on adjacent atoms of an aryl or heteroaryl ringin an aryl or heteroaryl group as described herein may optionally bereplaced with a substituent of the formula -T-C(O)—(CH₂)_(q)—U—, whereinT and U are independently —NH—, —O—, —CH₂— or a single bond, and q is aninteger of from 0 to 2. Alternatively, two of the substituents onadjacent atoms of the aryl or heteroaryl ring may optionally be replacedwith a substituent of the formula -J-(CH₂)_(r)—K—, wherein J and K areindependently —CH₂—, —O—, —NH—, —S—, —S(O)—, —S(O)₂—, —S(O)₂NR^(f)′— ora single bond, and r is an integer of from 1 to 3. One of the singlebonds of the new ring so formed may optionally be replaced with a doublebond. Alternatively, two of the substituents on adjacent atoms of thearyl or heteroaryl ring may optionally be replaced with a substituent ofthe formula —(CH₂)_(s)—X—(CH₂)_(t)—, where s and t are independentlyintegers of from 0 to 3, and X is —O—, —NR′—, —S—, —S(O)—, —S(O)₂—, or—S(O)₂NR^(a)′—. The substituent R^(f)′ in —NR^(f)′— and —S(O)₂NR^(f)′—is selected from hydrogen or unsubstituted (C₁-C₆) alkyl.

“Stable compound” and “stable structure” are meant to indicate acompound that is sufficiently robust to survive isolation to a usefuldegree of purity from a reaction mixture, and formulation into anefficacious therapeutic agent.

As used herein the term “therapeutically effective amount” generallymeans the amount necessary to ameliorate at least one symptom of adisorder to be prevented, reduced, or treated as described herein. Thephrase “therapeutically effective amount” as it relates to the GSNORinhibitors of the present invention shall mean the GSNOR inhibitordosage that provides the specific pharmacological response for which theGSNOR inhibitor is administered in a significant number of subjects inneed of such treatment. It is emphasized that a therapeuticallyeffective amount of a GSNOR inhibitor that is administered to aparticular subject in a particular instance will not always be effectivein treating the conditions/diseases described herein, even though suchdosage is deemed to be a therapeutically effective amount by those ofskill in the art.

C. S-Nitrosoglutathione Reductase Inhibitors

1. Inventive Compounds

In one of its aspects the present invention provides a compound having astructure shown in Formula I, or a pharmaceutically acceptable salt,stereoisomer, or prodrug thereof:

wherein:Ar is selected from the group consisting of phenyl and thiophen-yl;R₁ is selected from the group consisting of unsubstituted imidazolyl,substituted imidazolyl, chloro, bromo, fluoro, hydroxy, and methoxy;R₂ is selected from the group consisting of hydrogen, methyl, chloro,fluoro, hydroxy, methoxy, ethoxy, propoxy, carbamoyl, dimethylamino,amino, formamido, and trifluoromethyl; andX is selected from the group consisting of CO and SO₂.

In a further aspect of the invention, suitable identities for R₁include, but are not limited to, unsubstituted imidazolyl andsubstituted imidazolyl. Suitable substitutions for the substitutedimidazolyl group include, but are not limited to, C₁-C₆ alkyl.

In a further aspect of the invention ArR₁R₂ identities include, but arenot limited to,

wherein R₃ is selected from H, methyl, and ethyl.

In a further aspect of the invention, ArR₁ identities include, but arenot limited to, 4-chlorophenyl, 3-chlorophenyl, 4-bromophenyl,3-bromophenyl, 4-fluorophenyl, 3-fluorophenyl, 4-hydroxyphenyl,4-methoxyphenyl, 3-methoxyphenyl, 2-methoxyphenyl,4-chlorothiophen-2-yl, 5-chlorothiophen-2-yl, 3-bromothiophen-2-yl,4-bromothiophen-2-yl, 5-bromothiopheny-2-yl, and 5-bromothiophen-3-yl.

In one of its aspects the present invention provides a compound having astructure shown in Formula II, or a pharmaceutically acceptable salt,stereoisomer, or prodrug thereof:

wherein:Ar is selected from the group consisting of phenyl and thiophen-yl;R₄ is selected from the group consisting of unsubstituted imidazolyl andsubstituted imidazolyl;R₅ is selected from the group consisting of hydrogen, fluoro, hydroxy,and methoxy;R₆ is selected from the group consisting of hydrogen, chloro, bromo, andfluoro;R₇ is selected from the group consisting of hydrogen, and methyl; andR₈ is selected from the group consisting of CONH₂, SO₂NH₂, and NHSO₂CH₃.

In a further aspect of the invention, suitable identities for ArR₄R₅include, but are not limited to,

wherein R₉ is selected from H, methyl, and ethyl.

When a bond to a substituent is shown to cross a bond connecting twoatoms in a ring, then such substituent may be bonded to any atom in thering. When a substituent is listed without indicating the atom via whichsuch substituent is bonded to the rest of the compound of a givenformula, then such substituent may be bonded via any atom in suchsubstituent. Combinations of substituents and/or variables arepermissible, but only if such combinations result in stable compounds.

The compounds described herein may have asymmetric centers. Compounds ofthe present invention containing an asymmetrically substituted atom maybe isolated in optically active or racemic forms. It is well known inthe art how to prepare optically active forms, such as by resolution ofracemic forms or by synthesis from optically active starting materials.Many geometric isomers of olefins, C═N double bonds, and the like canalso be present in the compounds described herein, and all such stableisomers are contemplated in the present invention. Cis and transgeometric isomers of the compounds of the present invention aredescribed and may be isolated as a mixture of isomers or as separatedisomeric forms. All chiral, diastereomeric, racemic, and geometricisomeric forms of a structure are intended, unless the specificstereochemistry or isomeric form is specifically indicated. Alltautomers of shown or described compounds are also considered to be partof the present invention.

It is to be understood that isomers arising from such asymmetry (e.g.,all enantiomers and diastereomers) are included within the scope of theinvention, unless indicated otherwise. Such isomers can be obtained insubstantially pure form by classical separation techniques and bystereochemically controlled synthesis. Furthermore, the structures andother compounds and moieties discussed in this application also includeall tautomers thereof. Alkenes can include either the E- or Z-geometry,where appropriate.

2. Representative GSNOR Inhibitors

Table 1 below lists representative novel pyrrole analogs of Formula Iand Formula II useful as GSNOR inhibitors of the invention. Thesynthetic methods that can be used to prepare each compound, identifiedin Table 1 (i.e. Scheme 1, Scheme 2, etc.) are detailed below. In somecases, if the starting material or intermediate of a scheme is notcommercially available, then a corresponding method describes thesynthesis of that starting material or intermediate (i.e. Method 1,Method 2, etc.). Table 1 provides Scheme number, defines startingmaterials shown in the Schemes, and where necessary, provides a Methodnumber which corresponds to a detailed synthesis of an intermediate orstarting material. Supporting mass spectrometry data for each compoundis also included in Table 1. GSNOR inhibitor activity was determined bythe assay described in Example 2 and IC₅₀ values were obtained. GSNORinhibitor compounds 1-70 of Table 1 had an IC₅₀ of about <15 μM. GSNORinhibitor compounds 1-12, 14-15, 17-19, 22-36, 38-42, 44-56, 58-69 ofTable 1 had an IC₅₀ of about less than 1.0 μM.

TABLE 1 Mo- lecular Mass # Structure Compound name Chemical formulaweight Spec Scheme #/Method #  1

3-(5-(4-(1H-imidazol-1- yl)phenyl)-1-(4- carbamoyl-2- methylphenyl)-1H-pyrrol-2-yl)propanoic acid C24H22N4O3 414.5 415.1 Scheme 5, Ar1 = 4-carbamoyl-2- methylphenyl, R = H  2

3-(5-(5-(1H-imidazol-1- yl)thiophen-2-yl)-1-(4- carbamoyl-2-methylphenyl)-1H- pyrrol-2-yl)propanoic acid C22H20N4O3S 420.5 421.1Scheme 9b, Ar = 1H-imidazol-1- yl  3

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4-(2- methyl-1H-imidazol-1-yl)phenyl)-1H-pyrrol-2- yl)propanoic acid C25H24N4O3 428.5 429.1 Scheme9a, Ar = 2- methyl-1H-imidazol- 1-yl  4

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- hydroxyphenyl)-1H-pyrrol-2-yl)propanoic acid C21H20N2O4 364.4 365.1 Scheme 1, R2 = 4-hydroxyphenyl, R1 = 4-carbamoyl-2- methylphenyl  5

3-(5-(5-bromothiophen- 2-yl)-1-(4-carbamoyl-2- methylphenyl)-1H-pyrrol-2-yl)propanoic acid C19H17BrN2O3S 433.3 433,  435   Scheme 1, R2= 5- bromothiophen-2-yl, R1 = 4-carbamoyl-2- methylphenyl  6

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- methoxyphenyl)-1H-pyrrol-2-yl)propanoic acid C22H22N2O4 378.4 379.1 Scheme 1, R2 = 4-methoxyphenyl, R1 = 4-carbamoyl-2- methylphenyl  7

3-(5-(4-bromophenyl)-1- (4-carbamoyl-2- methylphenyl)-1H-pyrrol-2-yl)propanoic acid C21H19BrN2O3 427.3  427.1, 429.1 Scheme 6,Ar2 = 4- bromophenyl  8

3-(1-(4-carbamoyl-2- methylphenyl)-5-(3- chloro-4- methoxyphenyl)-1H-pyrrol-2-yl)propanoic acid C22H21ClN2O4 412.9 413.1 Scheme 6, Ar2 = 3-chloro-4- methoxyphenyl  9

3-(1-(4-carbamoyl-2- methylphenyl)-5-(3- fluoro-4- methoxyphenyl)-1H-pyrrol-2-yl)propanoic acid C22H21FN2O4 396.4 397.1 Scheme 6, Ar2 = 3-fluoro-4- methoxyphenyl 10

3-(1-(4-carbamoyl-2- methylphenyl)-5-(3- chloro-4- hydroxyphenyl)-1H-pyrrol-2-yl)propanoic acid C21H19ClN2O4 398.8 399.1 Scheme 1, R2 = 3-chloro-4- hydroxyphenyl, R1 = 4-carbamoyl-2- methylphenyl 11

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- methoxy-3- methylphenyl)-1H-pyrrol-2-yl)propanoic acid C23H24N2O4 392.4 393.2 Scheme 6, Ar2 = 4-methoxy-3- methylphenyl 12

3-(1-(4-carbamoyl-2- methylphenyl)-5-(3- methoxyphenyl)-1H-pyrrol-2-yl)propanoic acid C22H22N2O4 378.4 379.1 Scheme 1, R2 = 3-methoxyphenyl, R1 = 4-carbamoyl-2- methylphenyl 13

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4-(4- methyl-1H-imidazol-1-yl)phenyl)-1H-pyrrol-2- yl)propanoic acid C25H24N4O3 428.5 429.1 Scheme9a, Ar = 4- methyl-1H-imidazol- 1-yl 14

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4-(2- ethyl-1H-imidazol-1-yl)phenyl)-1H-pyrrol-2- yl)propanoic acid C26H26N4O3 442.5 443.2 Scheme5, Ar1 = 4- carbamoyl-2- methylphenyl, R = ethyl 15

3-(5-(4-amino-3- chlorophenyl)-1-(4- carbamoyl-2- methylphenyl)-1H-pyrrol-2-yl)propanoic acid C21H20ClN3O3 397.9 398.0 Scheme 6, Ar2 = 4-amino-3- chlorophenyl/method # 12 16

3-(1-(4-carbamoyl-2- methylphenyl)-5-(3,4- difluorophenyl)-1H-pyrrol-2-yl)propanoic acid C21H18F2N2O3 384.4 385.0 Scheme 6, Ar2 = 3,4-difluorophenyl 17

3-(1-(4-carbamoyl-2- methylphenyl)-5-(2,4- difluorophenyl)-1H-pyrrol-2-yl)propanoic acid C21H18F2N2O3 384.4 385.0 Scheme 6, Ar2 = 2,4-difluorophenyl 18

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- chlorophenyl)-1H-pyrrol-2-yl)propanoic acid C21H19ClN2O3 382.8 383.0 Scheme 6, Ar2 = 4-chlorophenyl 19

3-(5-(4-bromothiophen- 2-yl)-1-(4-carbamoyl-2- methylphenyl)-1H-pyrrol-2-yl)propanoic acid C19H17BrN2O3S 433.3  433.0, 434.8 Scheme 1,R2 = 4- bromothiophen-2-yl, R1 = 4-carbamoyl-2- methylphenyl 20

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- fluoro-3- methoxyphenyl)-1H-pyrrol-2-yl)propanoic acid C22H21FN2O4 396.4 397.2 Scheme 6, Ar2 = 4-fluoro-3- methoxyphenyl 21

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- carbamoyl-3-fluorophenyl)-1H-pyrrol- 2-yl)propanoic acid C22H2OFN3O4 409.4 410.2Scheme 6, Ar2 = 4- carbamoyl-3- fluorophenyl 22

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- methoxy-2- methylphenyl)-1H-pyrrol-2-yl)propanoic acid C23H24N2O4 392.4 393.2 Scheme 6, Ar2 = 4-methoxy-2- methylphenyl 23

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- chloro-2-fluorophenyl)-1H-pyrrol-2- yl)propanoic acid C21H18ClFN2O3 400.8 401.0 Scheme 6, Ar2 =4- chloro-2- fluorophenyl 24

3-(l-(4-carbamoyl-2- methylphenyl)-5-(4- fluorophenyl)-1H-pyrrol-2-yl)propanoic acid C21H19FN2O3 366.4 367.0 Scheme 6, Ar2 = 4-fluorophenyl 25

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- fluoro-2-methylphenyl)-1H-pyrrol-2- yl)propanoic acid C22H21FN2O3 380.4 381.1 Scheme 6, Ar2 =4- fluoro-2- methylphenyl 26

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- chloro-2- methoxyphenyl)-1H-pyrrol-2-yl)propanoic acid C22H21ClN2O4 412.9 413.0 Scheme 33, R1 = 4-carbamoyl-2- methylphenyl, R2 = 4-chloro, R3 = methyl 27

3-(1-(4-carbamoyl-2- methylphenyl)-5-(2- chloro-4- methoxyphenyl)-1H-pyrrol-2-yl)propanoic acid C22H21ClN2O4 412.9 414.0 Scheme 6, Ar2 = 2-chloro-4- methoxyphenyl 28

3-(5-(4-(1H-imidazol-1- yl)thiophen-2-yl)-1-(4- carbamoyl-2-methylphenyl)-1H- pyrrol-2-yl)propanoic acid C22H20N4O3S 420.5 421.1Followed procedure described in Scheme 9b, where starting material iscompound #19 in this table (before hydrolysis), Ar = 1H-imidazol-1- yl29

3-(1-(4-carbamoyl-2- methylphenyl)-5-(2- ethoxy-4-fluorophenyl)-1H-pyrrol-2- yl)propanoic acid C23H23FN2O4 410.4 411.2 Scheme 6, Ar2 =2- ethoxy-4- fluorophenyl 30

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- methoxy-2-(trifluoromethyl)phenyl)- 1H-pyrrol-2- yl)propanoic acid C23H21F3N2O4446.4 447.2 Scheme 6, Ar2 = 4- methoxy-2- (trifluoromethyl) phenyl 31

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- fluoro-2- methoxyphenyl)-1H-pyrrol-2-yl)propanoic acid C22H21FN2O4 396.4 397.1 Scheme 6, Ar2 = 4-fluoro-2- methoxyphenyl 32

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- chloro-3-fluorophenyl)-1H-pyrrol-2- yl)propanoic acid C21H18ClFN2O3 400.8 401.1 Scheme 6, Ar2 =4- chloro-3- fluorophenyl 33

3-(1-(4-carbamoyl-2- methylphenyl)-5-(5-(2- methyl-1H-imidazol-1-yl)thiophen-2-yl)-1H- pyrrol-2-yl)propanoic acid C23H22N4O3S 434.5 435.0Scheme 9b, Ar = 2- methyl-1H-imidazol- 1-yl 34

3-(1-(4-carbamoyl-2- methylphenyl)-5-(3- fluoro-4-(1H-imidazol-1-yl)phenyl)-1H-pyrrol-2- yl)propanoic acid C24H21FN4O3 432.4 433.1 Scheme36, R1 = 4- carbamoyl-2- methylphenyl, where 1st step followed alternateconditions of Scheme 36A, R = H 35

3-(1-(4-carbamoyl-2- methylphenyl)-5-(3- fluoro-4-(2-methyl-1H-imidazol-1-yl)phenyl)- 1H-pyrrol-2- yl)propanoic acid C25H23FN4O3 446.5447.1 Scheme 36, R1 = 4- carbamoyl-2- methylphenyl, where 1st stepfollowed alternate conditions of Scheme 36A, R = Me 36

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- chloro-2-ethoxyphenyl)-1H-pyrrol-2- yl)propanoic acid C23H23ClN2O4 426.9 427.1 Scheme 33, R1 =4- carbamoyl-2- methylphenyl, R2 = 4-chloro, R3 = ethyl 37

3-(5-(5-bromo-2- methoxyphenyl)-1-(4- carbamoyl-2- methylphenyl)-1H-pyrrol-2-yl)propanoic acid C22H21BrN2O4 457.3 459.0 Scheme 1, R1 = 4-carbamoyl-2- methylphenyl, R2 = 5-bromo-2- methoxyphenyl/ Method 41 38

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4-(2- methyl-1H-imidazol-1-yl)thiophen-2-yl)-1H- pyrrol-2-yl)propanoic acid C23H22N4O3S 434.5 435.2Followed procedure described in Scheme 9b, where starting material iscompound #19 in this table (before hydrolysis), , Ar = 2-methyl-1H-imidazol- 1-yl 39

3-(5-(4-bromo-2- methoxyphenyl)-1-(4- carbamoyl-2- methylphenyl)-1H-pyrrol-2-yl)propanoic acid C22H21BrN2O4 457.3 459.1 Scheme 1, R2 = 4-bromo-2- methoxyphenyl, R1 = 4-carbamoyl-2- methylphenyl/ Method 15 40

3-(1-(4-carbamoyl-2- methylphenyl)-5-(2- methoxy-4-(2-methyl-1H-imidazol-1- yl)phenyl)-1H-pyrrol-2- yl)propanoic acid C26H26N4O4458.5 459.1 Scheme 34, Ar1- X = 4-bromo-2- methoxyphenyl, Ar2 =2-methyl-1H- imidazol-1-yl/see previous compound for synthesis of 34A 41

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- chloro-2- hydroxyphenyl)-1H-pyrrol-2-yl)propanoic acid C21H19ClN2O4 398.8 399.0 Scheme 1, R1 = 4-carbamoyl-2- methylphenyl, R2 = 4-chloro-2- hydroxyphenyl 42

3-(5-(5-bromothiophen- 3-yl)-1-(4-carbamoyl-2- methylphenyl)-1H-pyrrol-2-yl)propanoic acid C19H17BrN2O3S 433.3 434.9 Scheme 1, R2 = 5-bromothiophen-3-yl, R1 = 4-carbamoyl-2- methylphenyl/ Method 19 43

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- hydroxy-3- methylphenyl)-1H-pyrrol-2-yl)propanoic acid C22H22N2O4 378.4 379.1 Scheme 1, R2 = 4-hydroxy-3- methylphenyl, R1 = 4-carbamoyl-2- methylphenyl 44

3-(1-(4-carbamoyl-2- methylphenyl)-5-(2- carbamoyl-4- chlorophenyl)-1H-pyrrol-2-yl)propanoic acid C22H20ClN3O4 425.9 426.1 Scheme 6, Ar2 = 2-carbamoyl-4- chlorophenyl, using 4-chloro-2- cyanophenylboronic acid instep 6E to 6F 45

3-(1-(4-carbamoyl-2- methylphenyl)-5-(2- methoxyphenyl)-1H-pyrrol-2-yl)propanoic acid C22H22N2O4 378.4 379.1 Scheme 6, Ar2 = 2-methoxyphenyl 46

3-(1-(4-carbamoyl-2- methylphenyl)-5-(2,4- dimethoxyphenyl)-1H-pyrrol-2-yl)propanoic acid C23H24N2O5 408.4 409.2 Scheme 19, Ar2 = 2,4-dimethoxyphenyl 47

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- chloro-2- propoxyphenyl)-1H-pyrrol-2-yl)propanoic acid C24H25ClN2O4 440.9 441.1 Scheme 33, R1 = 4-carbamoyl-2- methylphenyl, R2 = 4-chloro, R3 = n-propyl 48

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- hydroxy-2- methoxyphenyl)-1H-pyrrol-2-yl)propanoic acid C22H22N2O5 394.4 395.1 Scheme 6, R2 = 4-hydroxy-2- methoxyphenyl/ Method 18 49

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- chloro-2-(dimethylamino)phenyl)- 1H-pyrrol-2- yl)propanoic acid C23H24ClN3O3425.9 426.1 Scheme 39 50

3-(5-(4-(1H-imidazol-1- yl)-2-methoxyphenyl)-1- (4-carbamoyl-2-methylphenyl)-1H- pyrrol-2-yl)propanoic acid C25H24N4O4 444.5 445.2Scheme 36, R1 = 1H-imidazol-1- yl, R2 = 4- carbamoyl-2- methylphenyl,and R3 = 2- methoxy/method 15 51

3-(1-(4-carbamoyl-2- methylphenyl)-5-(5-(2- methyl-1H-imidazol-1-yl)thiophen-3-yl)-1H- pyrrol-2-yl)propanoic acid C23H22N4O3S 434.5 435.1Scheme 34, Ar1- X = 5- bromothiophen-3-yl, Ar2 = 2-methyl-1H-imidazol-1-yl, R1 = 4-carbamoyl-2- methylphenyl 52

3-(1-(4-carbamoyl-2- methylphenyl)-5-(5- chlorothiophen-2-yl)-1H-pyrrol-2- yl)propanoic acid C19H17ClN2O3S 388.9 389.0 Scheme 1, R2 =5- chlorothiophen-2-yl, R1 = 4-carbamoyl-2- methylphenyl 53

3-(1-(4-carbamoyl-2- methylphenyl)-5-(5-(2- ethyl-1H-imidazol-1-yl)thiophen-2-yl)-1H- pyrrol-2-yl)propanoic acid C24H24N4O3S 448.5 449.1Scheme 9b, Ar = 2- ethyl-1H-imidazol-1- yl 54

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- chloro-2- formamidophenyl)-1H-pyrrol-2-yl)propanoic acid C22H20ClN3O4 425.9 425.9 Scheme 40 55

3-(l-(4-carbamoyl-2- methylphenyl)-5-(3- chlorothiophen-2-yl)-1H-pyrrol-2- yl)propanoic acid C19H17ClN2O3S 388.9 389.0 Scheme 1, R1 =4- carbamoyl-2- methylphenyl, R2 = 3- chlorothiophen-2- yl/method 24 56

3-(1-(4-carbamoyl-2- methylphenyl)-5-(4- formamido-2- methoxyphenyl)-1H-pyrrol-2-yl)propanoic acid C23H23N3O5 421.4 422.0 Scheme 6, 4-formamido-2- methoxyphenyl/ Method 33 57

3-(5-(3-bromo-5- methoxythiophen-2-yl)- 1-(4-carbamoyl-2-methylphenyl)-1H- pyrrol-2-yl)propanoic acid C20H19BrN2O4S 463.3 464.6Scheme 1, R1 = 4- carbamoyl-2- methylphenyl, R2 = 3-bromo-5-methoxythiophen-2- yl/method 25 58

3-(l-(4-carbamoyl-2- methylphenyl)-5-(4- chlorothiophen-2-yl)-1H-pyrrol-2- yl)propanoic acid C19H17ClN2O3S 388.9 388.9 Scheme 1, R1 =4- carbamoyl-2- methylphenyl, R2 = 4- chlorothiophen-2- yl/Method 28(28-3) 59

3-(5-(5-bromo-4- chlorothiophen-2-yl)-1- (4-carbamoyl-2-methylphenyl)-1H- pyrrol-2-yl)propanoic acid C19H16BrClN2O3S 467.8 466.9, 468.8 Scheme 1, R1 = 4- carbamoyl-2- methylphenyl, R2 = 4-chlorothiophen-2- yl/Method 28 (28-2) 60

3-(5-(4-bromothiophen- 2-yl)-1-(2-methyl-4- sulfamoylphenyl)-1H-pyrrol-2-yl)propanoic acid C18H17BrN2O4S2 469.4 470.9 Scheme 1, R1 = 2-methyl-4- (methylsulfonamido) phenyl, R2 = 4- bromothiophen-2- yl/Method27 61

3-(5-(5-(2-methyl-1H- imidazol-1-yl)thiophen- 2-yl)-1-(2-methyl-4-sulfamoylphenyl)-1H- pyrrol-2-yl)propanoic acid C22H22N4O4S2 470.6 471.0Scheme 36, Ar1- Br = 5-bromo- thiophen-2-yl, Ar2 = 2-methyl-1H-imidazol-1-yl, R1 = 2-methyl-4- sulfamoylphenyl/ Method 27 62

3-(5-(5-(2-methyl-1H- imidazol-1-yl)thiophen- 2-yl)-1-(4-sulfamoylphenyl)-1H- pyrrol-2-yl)propanoic acid C21H20N4O4S2 456.5 457.0Scheme 34, Ar1- X = 5- bromothiophen-2-yl, Ar2 = 2-methyl-1H-imidazol-1-yl, R1 = 4- sulfamoylphenyl/ Method 35 63

3-(5-(5-(2-methyl-1H- imidazol-1-yl)thiophen- 2-yl)-1-(2-methyl-4-(methylsulfonamido) phenyl)-1H-pyrrol-2- yl)propanoic acidC23H23BrN4O4S2 563.5  563.0, 565.0 Prepared as a by- product duringsynthesis of compound # 67 of this table (formed in step 1 of Scheme 34,where it was isolated after hydrolysis by prep HPLC) 64

3-(5-(4-(1H-imidazol-1- yl)phenyl)-1-(2-methyl- 4-(methylsulfonamido)phenyl)-1H-pyrrol-2- yl)propanoic acid C24H24N4O4S 464.5 465.0 Scheme20, Ar2 = 4- (1H-imidazol-1- yl)phenyl/where 20A is compound # 1 in thistable 65

3-(5-(4-(2-methyl-1H- imidazol-1-yl)phenyl)-1- (2-methyl-4-(methylsulfonamido) phenyl)-1H-pyrrol-2- yl)propanoic acid C25H26N4O4S478.6 479.2 Scheme 20, Ar2 = 4- (2-methyl-1H- imidazol-1-yl)phenyl/where 20A is compound # 3 in this table 66

3-(5-(4-(2-methyl-1H- imidazol-1-yl)thiophen- 2-yl)-1-(2-methyl-4-(methylsulfonamido) phenyl)-1H-pyrrol-2- yl)propanoic acid C23H24N4O4S2484.6 485.1 Scheme 34, Ar1- X = 4- bromothiophen-2-yl, Ar2 =2-methyl-1H- imidazol-1-yl, R1 = 2-methyl-4- (methylsulfonamido)phenyl/Method 23 67

3-(5-(5-(2-methyl-1H- imidazol-1-yl)thiophen- 2-yl)-1-(2-methyl-4-(methylsulfonamido) phenyl)-1H-pyrrol-2- yl)propanoic acid C23H24N4O4S2484.6 485.0 Scheme 34, Ar1- X = 5- bromothiophen-2-yl, Ar2 =2-methyl-1H- imidazol-1-yl, R1 = 2-methyl-4- (methylsulfonamido)phenyl/Method 23 68

3-(5-(4-(2-methyl-1H- imidazol-1-yl)thiophen- 2-yl)-1-(4-(methylsulfonamido) phenyl)-1H-pyrrol-2- yl)propanoic acid C22H22N4O4S2470.6 470.9 Scheme 34, Ar1- X = 4- bromothiophen-2-yl, Ar2 =2-methyl-1H- imidazol-1-yl, R1 = 4- (methylsulfonamido) phenyl/Method 2369

3-(5-(2-methoxy-4-(2- methyl-1H-imidazol-1- yl)phenyl)-1-(4-(methylsulfonamido) phenyl)-1H-pyrrol-2- yl)propanoic acid C25H26N4O5S494.6 495.1 Scheme 34, Ar1- X = 2-methoxy-4- bromophenyl, Ar2 =2-methyl-1H- imidazol-1-yl, R1 = 4- (methylsulfonamido) phenyl/Method 23and Method 15 70

3-(5-(4-(2-methyl-1H- imidazol-1-yl)phenyl)-1- (4-(methylsulfonamido)phenyl)-1H-pyrrol-2- yl)propanoic acid C24H24N4O4S 464.5 464.9 Scheme34, Ar1- X = 4-bromophenyl, Ar2 = 2-methyl-1H- imidazol-1-yl, R1 = 4-(methylsulfonamido) phenyl/Method 23

D. Pharmaceutical Compositions Comprising a GSNOR Inhibitor

The invention encompasses pharmaceutical compositions comprising atleast one GSNOR inhibitor described herein and at least onepharmaceutically acceptable carrier. Suitable carriers are described in“Remington: The Science and Practice, Twentieth Edition,” published byLippincott Williams & Wilkins, which is incorporated herein byreference. Pharmaceutical compositions according to the invention mayalso comprise one or more non-GSNOR inhibitor active agents.

The pharmaceutical compositions of the invention can comprise novelGSNOR inhibitors described herein, the pharmaceutical compositions cancomprise known compounds which previously were not know to have GSNORinhibitor activity, or a combination thereof.

The GSNOR inhibitors can be utilized in any pharmaceutically acceptabledosage form, including but not limited to injectable dosage forms,liquid dispersions, gels, aerosols, ointments, creams, lyophilizedformulations, dry powders, tablets, capsules, controlled releaseformulations, fast melt formulations, delayed release formulations,extended release formulations, pulsatile release formulations, mixedimmediate release and controlled release formulations, etc.Specifically, the GSNOR inhibitors described herein can be formulated:(a) for administration selected from the group consisting of oral,pulmonary, intravenous, intra-arterial, intrathecal, intra-articular,rectal, ophthalmic, colonic, parenteral, intracisternal, intravaginal,intraperitoneal, local, buccal, nasal, and topical administration; (b)into a dosage form selected from the group consisting of liquiddispersions, gels, aerosols, ointments, creams, tablets, sachets andcapsules; (c) into a dosage form selected from the group consisting oflyophilized formulations, dry powders, fast melt formulations,controlled release formulations, delayed release formulations, extendedrelease formulations, pulsatile release formulations, and mixedimmediate release and controlled release formulations; or (d) anycombination thereof.

For respiratory infections, an inhalation formulation can be used toachieve high local concentrations. Formulations suitable for inhalationinclude dry powder or aerosolized or vaporized solutions, dispersions,or suspensions capable of being dispensed by an inhaler or nebulizerinto the endobronchial or nasal cavity of infected patients to treatupper and lower respiratory bacterial infections.

Solutions or suspensions used for parenteral, intradermal, orsubcutaneous application can comprise one or more of the followingcomponents: (1) a sterile diluent such as water for injection, salinesolution, fixed oils, polyethylene glycols, glycerine, propylene glycolor other synthetic solvents; (2) antibacterial agents such as benzylalcohol or methyl parabens; (3) antioxidants such as ascorbic acid orsodium bisulfite; (4) chelating agents such asethylenediaminetetraacetic acid; (5) buffers such as acetates, citratesor phosphates; and (5) agents for the adjustment of tonicity such assodium chloride or dextrose. The pH can be adjusted with acids or bases,such as hydrochloric acid or sodium hydroxide. A parenteral preparationcan be enclosed in ampoules, disposable syringes or multiple dose vialsmade of glass or plastic.

Pharmaceutical compositions suitable for injectable use may comprisesterile aqueous solutions (where water soluble) or dispersions andsterile powders for the extemporaneous preparation of sterile injectablesolutions or dispersion. For intravenous administration, suitablecarriers include physiological saline, bacteriostatic water, CremophorEL (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In allcases, the composition must be sterile and should be fluid to the extentthat easy syringability exists. The pharmaceutical composition should bestable under the conditions of manufacture and storage and should bepreserved against the contaminating action of microorganisms such asbacteria and fungi.

The carrier can be a solvent or dispersion medium comprising, forexample, water, ethanol, polyol (for example, glycerol, propyleneglycol, and liquid polyethylene glycol, and the like), and suitablemixtures thereof. The proper fluidity can be maintained, for example, bythe use of a coating such as lecithin, by the maintenance of therequired particle size in the case of dispersion and by the use ofsurfactants. Prevention of the action of microorganisms can be achievedby various antibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In manycases, it will be preferable to include isotonic agents, for example,sugars, polyalcohols such as manitol or sorbitol, and inorganic saltssuch as sodium chloride in the composition. Prolonged absorption of theinjectable compositions can be brought about by including in thecomposition an agent which delays absorption, for example, aluminummonostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating the activereagent (e.g., GSNOR inhibitor) in the required amount in an appropriatesolvent with one or a combination of ingredients enumerated above, asrequired, followed by filtered sterilization. Generally, dispersions areprepared by incorporating at least one GSNOR inhibitor into a sterilevehicle that contains a basic dispersion medium and any other requiredingredients. In the case of sterile powders for the preparation ofsterile injectable solutions, exemplary methods of preparation includevacuum drying and freeze-drying, both of which yield a powder of theGSNOR inhibitor plus any additional desired ingredient from a previouslysterile-filtered solution thereof.

Oral compositions generally include an inert diluent or an ediblecarrier. They can be enclosed, for example, in gelatin capsules orcompressed into tablets. For the purpose of oral therapeuticadministration, the GSNOR inhibitor can be incorporated with excipientsand used in the form of tablets, troches, or capsules. Oral compositionscan also be prepared using a fluid carrier for use as a mouthwash,wherein the compound in the fluid carrier is applied orally and swishedand expectorated or swallowed. Pharmaceutically compatible bindingagents, and/or adjuvant materials can be included as part of thecomposition.

For administration by inhalation, the compounds are delivered in theform of an aerosol spray from pressured container or dispenser thatcontains a suitable propellant, e.g., a gas such as carbon dioxide, anebulized liquid, or a dry powder from a suitable device. Fortransmucosal or transdermal administration, penetrants appropriate tothe barrier to be permeated are used in the formulation. Such penetrantsare generally known in the art, and include, for example, fortransmucosal administration, detergents, bile salts, and fusidic acidderivatives. Transmucosal administration can be accomplished through theuse of nasal sprays or suppositories. For transdermal administration,the active reagents are formulated into ointments, salves, gels, orcreams as generally known in the art. The reagents can also be preparedin the form of suppositories (e.g., with conventional suppository basessuch as cocoa butter and other glycerides) or retention enemas forrectal delivery.

In one embodiment, the GSNOR inhibitors are prepared with carriers thatwill protect against rapid elimination from the body. For example, acontrolled release formulation can be used, including implants andmicroencapsulated delivery systems. Biodegradable, biocompatiblepolymers can be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylactic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art.

Liposomal suspensions (including liposomes targeted to infected cellswith monoclonal antibodies to viral antigens) can also be used aspharmaceutically acceptable carriers. These can be prepared according tomethods known to those skilled in the art, for example, as described inU.S. Pat. No. 4,522,811.

Additionally, suspensions of the GSNOR inhibitors may be prepared asappropriate oily injection suspensions. Suitable lipophilic solvents orvehicles include fatty oils, such as sesame oil, or synthetic fatty acidesters, such as ethyl oleate, triglycerides, or liposomes. Non-lipidpolycationic amino polymers may also be used for delivery. Optionally,the suspension may also include suitable stabilizers or agents toincrease the solubility of the compounds and allow for the preparationof highly concentrated solutions.

It is especially advantageous to formulate oral or parenteralcompositions in dosage unit form for ease of administration anduniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the subject tobe treated; each unit containing a predetermined quantity of GSNORinhibitor calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier. The specificationfor the dosage unit forms of the invention are dictated by and directlydependent on the unique characteristics of the GSNOR inhibitor and theparticular therapeutic effect to be achieved, and the limitationsinherent in the art of compounding such an active agent for thetreatment of individuals.

Pharmaceutical compositions according to the invention comprising atleast one GSNOR inhibitor can comprise one or more pharmaceuticalexcipients. Examples of such excipients include, but are not limited tobinding agents, filling agents, lubricating agents, suspending agents,sweeteners, flavoring agents, preservatives, buffers, wetting agents,disintegrants, effervescent agents, and other excipients. Suchexcipients are known in the art. Exemplary excipients include: (1)binding agents which include various celluloses and cross-linkedpolyvinylpyrrolidone, microcrystalline cellulose, such as Avicel® PH101and Avicel® PH102, silicified microcrystalline cellulose (ProSolvSMCC™), gum tragacanth and gelatin; (2) filling agents such as variousstarches, lactose, lactose monohydrate, and lactose anhydrous; (3)disintegrating agents such as alginic acid, Primogel, corn starch,lightly crosslinked polyvinyl pyrrolidone, potato starch, maize starch,and modified starches, croscarmellose sodium, cross-povidone, sodiumstarch glycolate, and mixtures thereof; (4) lubricants, including agentsthat act on the flowability of a powder to be compressed, includemagnesium stearate, colloidal silicon dioxide, such as Aerosil® 200,talc, stearic acid, calcium stearate, and silica gel; (5) glidants suchas colloidal silicon dioxide; (6) preservatives, such as potassiumsorbate, methylparaben, propylparaben, benzoic acid and its salts, otheresters of parahydroxybenzoic acid such as butylparaben, alcohols such asethyl or benzyl alcohol, phenolic compounds such as phenol, orquaternary compounds such as benzalkonium chloride; (7) diluents such aspharmaceutically acceptable inert fillers, such as microcrystallinecellulose, lactose, dibasic calcium phosphate, saccharides, and/ormixtures of any of the foregoing; examples of diluents includemicrocrystalline cellulose, such as Avicel® PH101 and Avicel® PH102;lactose such as lactose monohydrate, lactose anhydrous, and Pharmatose®DCL21; dibasic calcium phosphate such as Emcompress®; mannitol; starch;sorbitol; sucrose; and glucose; (8) sweetening agents, including anynatural or artificial sweetener, such as sucrose, saccharin sucrose,xylitol, sodium saccharin, cyclamate, aspartame, and acesulfame; (9)flavoring agents, such as peppermint, methyl salicylate, orangeflavoring, Magnasweet® (trademark of MAFCO), bubble gum flavor, fruitflavors, and the like; and (10) effervescent agents, includingeffervescent couples such as an organic acid and a carbonate orbicarbonate. Suitable organic acids include, for example, citric,tartaric, malic, fumaric, adipic, succinic, and alginic acids andanhydrides and acid salts. Suitable carbonates and bicarbonates include,for example, sodium carbonate, sodium bicarbonate, potassium carbonate,potassium bicarbonate, magnesium carbonate, sodium glycine carbonate,L-lysine carbonate, and arginine carbonate. Alternatively, only thesodium bicarbonate component of the effervescent couple may be present.

E. Kits Comprising the Compositions of the Invention

The present invention also encompasses kits comprising the compositionsof the invention. Such kits can comprise, for example, (1) at least oneGSNOR inhibitor; and (2) at least one pharmaceutically acceptablecarrier, such as a solvent or solution. Additional kit components canoptionally include, for example: (1) any of the pharmaceuticallyacceptable excipients identified herein, such as stabilizers, buffers,etc., (2) at least one container, vial or similar apparatus for holdingand/or mixing the kit components; and (3) delivery apparatus, such as aninhaler, nebulizer, syringe, etc.

F. Methods of Preparing GSNOR Inhibitors

The GSNOR inhibitors of the invention can readily be synthesized usingknown synthetic methodologies or via a modification of known syntheticmethodologies. As would be readily recognized by a skilled artisan, themethodologies described below allow the synthesis of pyrroles having avariety of substituents. Exemplary synthetic methods are described inthe examples below.

According to one synthetic protocol, reaction of 2-furaldehyde with anappropriately substituted acetophenone followed by treatment with astrong acid gives the appropriately substituted 1,4,7-trione.Cyclization of the trione to the corresponding 1,2,5-trisubstitutedpyrrole is readily achieved by reacting the trione with a primary aminein the presence of p-toluenesulfonic acid. In one embodiment of thepresent invention, further derivatization of the phenyl ring at C5 ofthe pyrrole is readily achieved, for example, by various cross-couplingreactions. For example, synthesis of the trisubstituted pyrroles byreacting 1-(4-chlorophenyl) ethanone and 2-furaldehyde will give thetarget pyrrole with 4-chlorophenyl group at C5. The aryl chloride can bederivatized by reaction with a boronic acid under Suzuki couplingconditions. Such routine derivatization methodologies allow the rapidgeneration of compound libraries for in vitro GSNOR inhibition studies.A variety of additional methods are described in Example 1 of thisdocument.

If needed, further purification and separation of enantiomers anddiastereomers can be achieved by routine procedures known in the art.Thus, for example, the separation of enantiomers of a compound can beachieved by the use of chiral HPLC and related chromatographictechniques. Diastereomers can be similarly separated. In some instances,however, diastereomers can simply be separated physically, such as, forexample, by controlled precipitation or crystallization.

The process of the invention, when carried out as prescribed herein, canbe conveniently performed at temperatures that are routinely accessiblein the art. In one embodiment, the process is performed at a temperaturein the range of about 25° C. to about 110° C. In another embodiment, thetemperature is in the range of about 40° C. to about 100° C. In yetanother embodiment, the temperature is in the range of about 50° C. toabout 95° C.

Synthetic steps that require a base are carried out using any convenientorganic or inorganic base. Typically, the base is not nucleophilic.Thus, in one embodiment, the base is selected from carbonates,phosphates, hydroxides, alkoxides, salts of disilazanes, and tertiaryamines.

The process of the invention, when performed as described herein, can besubstantially complete after several minutes to after several hoursdepending upon the nature and quantity of reactants and reactiontemperature. The determination of when the reaction is substantiallycomplete can be conveniently evaluated by ordinary techniques known inthe art such as, for example, HPLC, LCMS, TLC, and ¹H NMR.

G. Method of Treatment

The invention encompasses methods of preventing or treating (e.g.,alleviating one or more symptoms of) medical conditions through use ofone or more of the disclosed compounds. The methods compriseadministering a therapeutically effective amount of a GSNOR inhibitor toa patient in need. The compositions of the invention can also be usedfor prophylactic therapy.

The GSNOR inhibitor used in the methods of treatment according to theinvention can be: (1) a novel GSNOR inhibitor described herein, or apharmaceutically acceptable salt thereof, a prodrug thereof, or ametabolite thereof; (2) a compound which was known prior to the presentinvention, but wherein it was not known that the compound is a GSNORinhibitor, or a pharmaceutically acceptable salt thereof, a prodrugthereof, or a metabolite thereof; or (3) a compound which was knownprior to the present invention, and wherein it was known that thecompound is a GSNOR inhibitor, but wherein it was not known that thecompound is useful for the methods of treatment described herein, or apharmaceutically acceptable salt thereof, a prodrug thereof, or ametabolite thereof.

The patient can be any animal, domestic, livestock or wild, including,but not limited to cats, dogs, horses, pigs and cattle, and preferablyhuman patients. As used herein, the terms patient and subject may beused interchangeably.

In subjects with deleteriously high levels of GSNOR or GSNOR activity,modulation may be achieved, for example, by administering one or more ofthe disclosed compounds that disrupts or down-regulates GSNOR function,or decreases GSNOR levels. These compounds may be administered withother GSNOR inhibitor agents, such as anti-GSNOR antibodies or antibodyfragments, GSNOR antisense, iRNA, or small molecules, or otherinhibitors, alone or in combination with other agents as described indetail herein.

The present invention provides a method of treating a subject afflictedwith a disorder ameliorated by NO donor therapy. Such a method comprisesadministering to a subject a therapeutically effective amount of a GSNORinhibitor.

As used herein, “treating” describes the management and care of apatient for the purpose of combating a disease, condition, or disorderand includes the administration of a compound of the present inventionto prevent the onset of the symptoms or complications, alleviating thesymptoms or complications, or eliminating the disease, condition ordisorder. More specifically, “treating” includes reversing, attenuating,alleviating, minimizing, suppressing or halting at least one deleterioussymptom or effect of a disease (disorder) state, disease progression,disease causative agent (e.g., bacteria or viruses), or other abnormalcondition. Treatment is continued as long as symptoms and/or pathologyameliorate.

The disorders can include pulmonary disorders associated with hypoxemiaand/or smooth muscle constriction in the lungs and/or lung infectionand/or lung injury (e.g., pulmonary hypertension, ARDS, asthma,pneumonia, pulmonary fibrosis/interstitial lung diseases, cysticfibrosis, COPD) cardiovascular disease and heart disease, includingconditions such as hypertension, ischemic coronary syndromes,atherosclerosis, glaucoma, diseases characterized by angiogenesis (e.g.,coronary artery disease), disorders where there is risk of thrombosisoccurring, disorders where there is risk of restenosis occurring,chronic inflammatory diseases (e.g., AID dementia and psoriasis),diseases where there is risk of apoptosis occurring (e.g., heartfailure, atherosclerosis, heart failure, degenerative neurologicdisorders, arthritis and liver injury (ischemic or alcoholic)),impotence, obesity caused by eating in response to craving for food,stroke, reperfusion injury (e.g., traumatic muscle injury in heart orlung or crush injury), and disorders where preconditioning of heart orbrain for NO protection against subsequent ischemic events isbeneficial.

In one embodiment, the compounds of the present invention or apharmaceutically acceptable salt thereof, or a prodrug or metabolitethereof, can be administered in combination with an NO donor. An NOdonor donates nitric oxide or a related redox species and more generallyprovides nitric oxide bioactivity, that is activity which is identifiedwith nitric oxide, e.g., vasorelaxation or stimulation or inhibition ofa receptor protein, e.g., ras protein, adrenergic receptor, NFκB. NOdonors including S-nitroso, O-nitroso, C-nitroso and N-nitroso compoundsand nitro derivatives thereof and metal NO complexes, but not excludingother NO bioactivity generating compounds, useful herein are describedin “Methods in Nitric Oxide Research,” Feelisch et al. eds., pages71-115 (J. S., John Wiley & Sons, New York, 1996), which is incorporatedherein by reference. NO donors which are C-nitroso compounds wherenitroso is attached to a tertiary carbon which are useful herein includethose described in U.S. Pat. No. 6,359,182 and in WO 02/34705. Examplesof S-nitroso compounds, including S-nitrosothiols useful herein,include, for example, S-nitrosoglutathione,S-nitroso-N-acetylpenicillamine, S-nitroso-cysteine and ethyl esterthereof, S-nitroso cysteinyl glycine,S-nitroso-gamma-methyl-L-homocysteine, S-nitroso-L-homocysteine,S-nitroso-gamma-thio-L-leucine, S-nitroso-delta-thio-L-leucine, andS-nitrosoalbumin. Examples of other NO donors useful herein are sodiumnitroprusside (nipride), ethyl nitrite, isosorbide, nitroglycerin, SIN 1which is molsidomine, furoxamines, N-hydroxy (N-nitrosamine) andperfluorocarbons that have been saturated with NO or a hydrophobic NOdonor.

The combination of a GSNOR inhibitor with R(+) enantiomer of amlodipine,a known NO releaser (Zhang X. P at al. 2002 J. CardiovascularPharmacology 39, 208-214) is also an embodiment of the presentinvention.

The present invention also provides a method of treating a subjectafflicted with pathologically proliferating cells where the methodcomprises administering to said subject a therapeutically effectiveamount of an inhibitor of GSNOR. The inhibitors of GSNOR are thecompounds as defined above, or a pharmaceutically acceptable saltthereof, or a prodrug or metabolite thereof, in combination with apharmaceutically acceptable carrier. Treatment is continued as long assymptoms and/or pathology ameliorate.

In another embodiment, the pathologically proliferating cells can bepathologically proliferating microbes. The microbes involved can bethose where GSNOR is expressed to protect the microbe from nitrosativestress or where a host cell infected with the microbe expresses theenzyme, thereby protecting the microbe from nitrosative stress. The term“pathologically proliferating microbes” is used herein to meanpathologic microorganisms including but not limited to pathologicbacteria, pathologic viruses, pathologic Chlamydia, pathologic protozoa,pathologic Rickettsia, pathologic fungi, and pathologic mycoplasmata.More detail on the applicable microbes is set forth at columns 11 and 12of U.S. Pat. No. 6,057,367. The term “host cells infected withpathologic microbes” includes not only mammalian cells infected withpathologic viruses but also mammalian cells containing intracellularbacteria or protozoa, e.g., macrophages containing Mycobacteriumtuberculosis, Mycobacterium leper (leprosy), or Salmonella typhi(typhoid fever).

In another embodiment, the pathologically proliferating cells can bepathologic helminths. The term “pathologic helminths” is used herein torefer to pathologic nematodes, pathologic trematodes and pathologiccestodes. More detail on the applicable helminths is set forth at column12 of U.S. Pat. No. 6,057,367.

In another embodiment, the pathologically proliferating cells can bepathologically proliferating mammalian cells. The term “pathologicallyproliferating mammalian cells” as used herein means cells of the mammalthat grow in size or number in said mammal so as to cause a deleteriouseffect in the mammal or its organs. The term includes, for example, thepathologically proliferating or enlarging cells causing restenosis, thepathologically proliferating or enlarging cells causing benign prostatichypertrophy, the pathologically proliferating cells causing myocardialhypertrophy and proliferating cells at inflammatory sites such assynovial cells in arthritis or cells associated with a cellproliferation disorder.

As used herein, the term “cell proliferative disorder” refers toconditions in which the unregulated and/or abnormal growth of cells canlead to the development of an unwanted condition or disease, which canbe cancerous or non-cancerous, for example a psoriatic condition. Asused herein, the term “psoriatic condition” refers to disordersinvolving keratinocyte hyperproliferation, inflammatory cellinfiltration, and cytokine alteration. The cell proliferative disordercan be a precancerous condition or cancer. The cancer can be primarycancer or metastatic cancer, or both.

As used herein, the term “cancer” includes solid tumors, such as lung,breast, colon, ovarian, pancreas, prostate, adenocarcinoma, squamouscarcinoma, sarcoma, malignant glioma, leiomyosarcoma, hepatoma, head andneck cancer, malignant melanoma, non-melanoma skin cancers, as well ashematologic tumors and/or malignancies, such as leukemia, childhoodleukemia and lymphomas, multiple myeloma, Hodgkin's disease, lymphomasof lymphocytic and cutaneous origin, acute and chronic leukemia such asacute lymphoblastic, acute myelocytic or chronic myelocytic leukemia,plasma cell neoplasm, lymphoid neoplasm and cancers associated withAIDS.

In addition to psoriatic conditions, the types of proliferative diseaseswhich may be treated using the compositions of the present invention areepidermic and dermoid cysts, lipomas, adenomas, capillary and cutaneoushemangiomas, lymphangiomas, nevi lesions, teratomas, nephromas,myofibromatosis, osteoplastic tumors, and other dysplastic masses andthe like. In one embodiment, proliferative diseases include dysplasiasand disorders of the like.

In one embodiment, the treating cancer comprises a reduction in tumorsize, decrease in tumor number, a delay of tumor growth, decrease inmetastaic lesions in other tissues or organs distant from the primarytumor site, an improvement in the survival of patients, or animprovement in the quality of patient life, or at least two of theabove.

In another embodiment, the treating a cell proliferative disordercomprises a reduction in the rate of cellular proliferation, reductionin the proportion of proliferating cells, a decrease in size of an areaor zone of cellular proliferation, or a decrease in the number orproportion of cells having an abnormal appearance or morphology, or atleast two of the above.

In yet another embodiment, the compounds of the present invention or apharmaceutically acceptable salt thereof, a prodrug thereof, ormetabolite thereof, can be administered in combination with a secondchemotherapeutic agent. In a further embodiment, the secondchemotherapeutic agent is selected from the group consisting oftamoxifen, raloxifene, anastrozole, exemestane, letrozole, cisplatin,carboplatin, paclitaxel, cyclophosphamide, lovastatin, minosine,gemcitabine, araC, 5-fluorouracil, methotrexate, docetaxel, goserelin,vincristin, vinblastin, nocodazole, teniposide, etoposide, epothilone,navelbine, camptothecin, daunonibicin, dactinomycin, mitoxantrone,amsacrine, doxorubicin, epirubicin, idarubicin imatanib, gefitinib,erlotinib, sorafenib, sunitinib malate, trastuzumab, rituximab,cetuximab, and bevacizumab.

In one embodiment, the compounds of the present invention or apharmaceutically acceptable salt thereof, a prodrug thereof, ormetabolite thereof, can be administered in combination with an agentthat imposes nitrosative or oxidative stress. Agents for selectivelyimposing nitrosative stress to inhibit proliferation of pathologicallyproliferating cells in combination therapy with GSNOR inhibitors hereinand dosages and routes of administration therefore include thosedisclosed in U.S. Pat. No. 6,057,367, which is incorporated herein.Supplemental agents for imposing oxidative stress (i.e., agents thatincrease GSSG (oxidized glutathione) over GSH (glutathione) ratio orNAD(P) over NAD(P)H ratio or increase thiobarbituric acid derivatives)in combination therapy with GS-FDH inhibitors herein include, forexample, L-buthionine-S-sulfoximine (BSO), glutathione reductaseinhibitors (e.g., BCNU), inhibitors or uncouplers of mitochondrialrespiration and drugs that increase reactive oxygen species (ROS), e.g.,adriamycin, in standard dosages with standard routes of administration.

GSNOR inhibitors may also be co-administered with a phosphodiesteraseinhibitor (e.g., rolipram, cilomilast, roflumilast, Viagra® (sildenifilcitrate), Cialis® (tadalafil), Levitra® (vardenifil), etc.), aβ-agonist, a steroid, or a leukotriene antagonist (LTD4). Those skilledin the art can readily determine the appropriate therapeuticallyeffective amount depending on the disorder to be ameliorated.

GSNOR inhibitors may be used as a means to improve β-adrenergicsignaling. In particular, inhibitors of GSNOR alone or in combinationwith β-agonists could be used to treat or protect against heart failure,or other vascular disorders such as hypertension and asthma. GSNORinhibitors can also be used to modulate G protein coupled receptors(GPCRs) by potentiating Gs G-protein, leading to smooth musclerelaxation (e.g., airway and blood vessels), and by attenuating GqG-protein, and thereby preventing smooth muscle contraction (e.g., inairway and blood vessels).

The therapeutically effective amount for the treatment of a subjectafflicted with a disorder ameliorated by NO donor therapy is the GSNORinhibiting amount in vivo that causes amelioration of the disorder beingtreated or protects against a risk associated with the disorder. Forexample, for asthma, a therapeutically effective amount is abronchodilating effective amount; for cystic fibrosis, a therapeuticallyeffective amount is an airway obstruction ameliorating effective amount;for ARDS, a therapeutically effective amount is a hypoxemia amelioratingeffective amount; for heart disease, a therapeutically effective amountis an angina relieving or angiogenesis inducing effective amount; forhypertension, a therapeutically effective amount is a blood pressurereducing effective amount; for ischemic coronary disorders, atherapeutic amount is a blood flow increasing effective amount; foratherosclerosis, a therapeutically effective amount is an endothelialdysfunction reversing effective amount; for glaucoma, a therapeuticamount is an intraocular pressure reducing effective amount; fordiseases characterized by angiogenesis, a therapeutically effectiveamount is an angiogenesis inhibiting effective amount; for disorderswhere there is risk of thrombosis occurring, a therapeutically effectiveamount is a thrombosis preventing effective amount; for disorders wherethere is risk of restenosis occurring, a therapeutically effectiveamount is a restenosis inhibiting effective amount; for chronicinflammatory diseases, a therapeutically effective amount is aninflammation reducing effective amount; for disorders where there isrisk of apoptosis occurring, a therapeutically effective amount is anapoptosis preventing effective amount; for impotence, a therapeuticallyeffective is an erection attaining or sustaining effective amount; forobesity, a therapeutically effective amount is a satiety causingeffective amount; for stroke, a therapeutically effective amount is ablood flow increasing or a TIA protecting effective amount; forreperfusion injury, a therapeutically effective amount is a functionincreasing effective amount; and for preconditioning of heart and brain,a therapeutically effective amount is a cell protective effectiveamount, e.g., as measured by triponin or CPK.

The therapeutically effective amount for the treatment of a subjectafflicted with pathologically proliferating cells means a GSNORinhibiting amount in vivo which is an antiproliferative effectiveamount. Such antiproliferative effective amount as used herein means anamount causing reduction in rate of proliferation of at least about 20%,at least about 10%, at least about 5%, or at least about 1%.

In general, the dosage, i.e., the therapeutically effective amount,ranges from 1 μg to 10 g/kg and often ranges from 10 μg to 1 g/kg or 10μg to 100 mg/kg body weight of the subject being treated, per day.

H. Other Uses

The compounds of the present invention or a pharmaceutically acceptablesalt thereof, or a prodrug or metabolite thereof, can be applied tovarious apparatus in circumstances when the presence of such compoundswould be beneficial. Such apparatus can be any device or container, forexample, implantable devices in which a GSNOR inhibitor can be used tocoat a surgical mesh or cardiovascular stent prior to implantation in apatient. The GSNOR inhibitors of the present invention can also beapplied to various apparatus for in vitro assay purposes or forculturing cells.

The compounds of the present invention or a pharmaceutically acceptablesalt thereof, or a prodrug or metabolite thereof, can also be used as anagent for the development, isolation or purification of binding partnersto GSNOR inhibitor compounds, such as antibodies, natural ligands, andthe like. Those skilled in the art can readily determine related usesfor the compounds of the present invention.

EXAMPLES

The following examples are given to illustrate the present invention. Itshould be understood, however, that the invention is not to be limitedto the specific conditions or details described in these examples.Throughout the specification, any and all references to a publiclyavailable document, including a U.S. patent, are specificallyincorporated by reference.

Example 1: General and Specific Methods of Preparing Novel GSNOR PyrroleInhibitors

This example describes schemes for preparing the GSNOR inhibitorsdepicted in Table 1. Some schemes are specific to a particular compound,while others are general schemes that include an exemplary method forpreparing a representative compound. Following the schemes are methodswhich describe the preparation of intermediates that were used in selectschemes.

Representative Procedure for Scheme 1: Synthesis of3-[1-(4-Carbamoyl-2-methyl-phenyl)-5-(4-methoxy-phenyl)-1H-pyrrol-2-yl]-propanoicacid Step 1: Synthesis of(E)-3-Furan-2-yl-1-(4-methoxy-phenyl)-propenone

A solution of 2-furaldehyde (5.85 g, 60.92 mmol) was added to a methanolsolution (120 mL) of 4-methoxy acetophenone (8.5 g, 56.6 mmol), followedby the addition of sodium methoxide (3.1 g, 56.6 mmol). The reactionmixture was stirred at room temperature for 16 h, followed by removal ofthe solvent in vacuo. The resultant mixture was diluted with water (130mL) and extracted with ethyl acetate (350 mL). The aqueous layer wasre-extracted with ethyl acetate (100 mL). The combined organic layerswere dried over anhydrous Na₂SO₄ and the solvent was removed in vacuo toobtain the product (E)-3-Furan-2-yl-1-(4-methoxy-phenyl)-propenone as anorange solid (12.6 g, 97%).

Step 2: Synthesis of 1-(4-Methoxy-phenyl)-decane-1,4,7-trione

Conc. HCl (59 mL) was added to a solution of(E)-3-Furan-2-yl-1-(4-methoxy-phenyl)-propenone (12.6 g, 55.2 mmol) inethanol (237 mL). The reaction mixture was heated under reflux for 16 h,concentrated, and diluted with dichloromethane (250 mL), and theresultant organic layer was washed with water (25 mL). After phaseseparation, the organic layer was dried over anhydrous Na₂SO₄ and thesolvent removed in vacuo to obtain the crude mixture, which was purifiedby silica gel flash chromatography to obtain1-(4-methoxy-phenyl)-decane-1,4,7-trione (6.89 g, 43%).

Step 3: Synthesis of3-[1-(4-Carbamoyl-2-methyl-phenyl)-5-(4-methoxy-phenyl)-1H-pyrrol-2-yl]propanoicacid ethyl ester

4-amino-3-methylbenzamide (180 mg, 1.2 mmol) was added to a solution1-(4-methoxy-phenyl)-decane-1,4,7-trione (350 mg, 1.2 mmol) in ethanol(6 mL), followed by the addition of p-toluenesulfonic acid monohydrate(abbreviated TsOH or pTsOH) (23 mg, 0.12 mmol). The reaction mixture washeated under reflux for 16 h, and the solvent removed in vacuo to obtaina crude product which upon purification by silica gel flashchromatography, gives3-[1-(4-carbamoyl-2-methyl-phenyl)-5-(4-methoxy-phenyl)-1H-pyrrol-2-yl]propanoicacid ethyl ester (147 mg, 30%).

Step 4: Synthesis of3-[1-(4-Carbamoyl-2-methyl-phenyl)-5-(4-methoxy-phenyl)-1H-pyrrol-2-yl]-propanoicacid

3-[1-(4-carbamoyl-2-methyl-phenyl)-5-(4-methoxy-phenyl)-1H-pyrrol-2-yl]propanoicacid ethyl ester (86 mg, 0.216 mmol) was dissolved in ethanol (4 mL).Water (0.5 mL) was added to the ethanolic solution followed by theaddition of 1N NaOH (0.51 mL, 0.51 mmol). The reaction mixture wasstirred at room temperature for 1 h and then at 45° C. for an additionalhour. After removal of the solvent in vacuo, the residue was dilutedwith water (6 mL) and extracted with ethyl acetate (2×6 mL). The pH ofthe aqueous layer was adjusted to 2 with 1N HCl and then extracted withethyl acetate (6 mL). The combined organic layer was dried overanhydrous Na₂SO₄ and the solvent removed in vacuo to obtain3-[1-(4-Carbamoyl-2-methyl-phenyl)-5-(4-methoxy-phenyl)-1H-pyrrol-2-yl]-propanoicacid as the product (68 mg, 85%).

Representative Procedure for Scheme 1A, Alternate Conditions: Synthesisof3-[1-(4-Carbamoyl-thiazol-2-yl)-5-(4-methoxy-phenyl)-1H-pyrrol-2-yl]-propionicacid Step 3: Synthesis of3-[1-(4-Carbamoyl-thiazol-2-yl)-5-(4-methoxyphpenyl)-1H-pyrrol-2-yl]-propionicacid ethyl ester (1C, R1=4-carbamoyl-thiazol-2-yl, R2=4-methoxy-phenyl)

To a solution of 7-(4-methoxy-phenyl)-4,7-dioxo-heptanoic acid ethylester (0.5 mmol), see scheme 1, in ethanol (2 mL) were added the amine(1.5 equivalents) and p-toluenesulfonic acid monohydrate (0.5 eq.). Thereaction was run using the Biotage Microwave Initiator for 1 to 3 hoursat 150° C. The solvent was removed in vacuo to obtain the crude mixturewhich was purified by prep silica gel plate to obtain the final product(70 mg, 38%).

Step 4: Synthesis of3-[1-(4-Carbamoyl-thiazol-2-yl)-5-(4-methoxyphpenyl)-1H-pyrrol-2-yl]-propionicacid (1D, R1=4-carbamoyl-thiazol-2-yl, R2=4-methoxy-phenyl)

To3-[1-(4-Carbamoyl-thiazol-2-yl)-5-(4-methoxy-phenyl)-1H-pyrrol-2-yl]-propionicacid ethyl ester (0.15 mmol) in a 2:1 mixture of methanol/THF was added2M LiOH (0.30 mmol). The reaction mixture was stirred for 24 hours. Thesolvent was removed in vacuo. The residue was diluted with water (2 mL)and extracted with ethyl ether. The pH of the aqueous layer was adjustedto 2 with 1N HCl. The resulting suspension was filtered; the solid waswashed with water and dried to give the final compound. Yield: 36 mg,69%.

Scheme 2-Scheme 4 intentionally omitted.

Representative Procedure for Scheme 5: Synthesis of3-(5-(4-(1H-imidazol-1-yl)phenyl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid (5E, Ar1=4-carbamoyl-2-methylphenyl, R═H) Step 1: Synthesis of1-(4-bromophenyl)-3-(furan-2-yl)prop-2-en-1-one (5A)

To a solution of 4-bromophenylethanone (112.6 g, 570 mmol) andfuran-2-carbaldehyde (58.5 g, 610 mmol) in methanol (1.5 L) was addedCH₃ONa (31 g, 570 mmol) over 10 min and the reaction solution wasstirred at room temperature overnight. The reaction mixture wasneutralized with conc. HCl to pH=7, and the solvent was removed underreduced pressure. To the resultant residue was added EA and water. Theaqueous layer was extracted with EA for 3 times. The combined layerswere washed with brine, dried over MgSO₄, concentrated and purified bysilica gel column chromatography (PE (petroleum ether): EA (ethylacetate)=10:1) to afford 1-(4-bromophenyl)-3-(furan-2-yl)prop-2-en-1-one(5A) as a yellow solid (90.2 g, 65%).

Step 2: Synthesis of ethyl 7-(4-bromophenyl)-4,7-dioxoheptanoate (5B)

To a solution of compound1-(4-bromophenyl)-3-(furan-2-yl)prop-2-en-1-one (5A) (20.0 g, 72.2 mmol)in ethanol (160 mL) was added HBr (48% in water, 40 mL). The resultantmixture was stirred under reflux for 8 h, and then the reaction solutionwas concentrated in vacuo. To the residue was added sat. NaHCO₃ to pH=7and extracted with EA. The combined organic layers were washed withbrine, dried over MgSO₄, concentrated and purified by silica gel columnchromatography (PE:EA=5:1) to afford ethyl7-(4-bromophenyl)-4,7-dioxoheptanoate (5B) as a yellow solid (7.0 g,28%).

Step 3: Synthesis of Ethyl3-(5-(4-bromophenyl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoate(5C, Ar1=4-carbamoyl-2-methylphenyl)

To a solution of ethyl 7-(4-bromophenyl)-4,7-dioxoheptanoate (5B) (3.41g, 10 mmol) and 4-amino-3-methylbenzamide (1.65 g, 11 mmol) in 50 mL ofethanol was added TsOH.H₂O (570 mg, 3 mmol). The reaction solution wasstirred under reflux overnight and then concentrated in vacuo. Theresultant residue was neutralized with sat. NaHCO₃ and extracted withEthyl Acetate. The organic layers were washed with brine, concentratedand purified by silica gel column chromatography (DCM:PE=1:1) to affordethyl 7-(4-bromophenyl)-4,7-dioxoheptanoate as a pale solid (2.80 g,61%).

Step 4: Synthesis of ethyl3-(5-(4-(1H-imidazol-1-yl)phenyl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoate(5D, Ar1=4-carbamoyl-2-methylphenyl, R═H)

To a mixture of ethyl 7-(4-bromophenyl)-4,7-dioxoheptanoate (4.54 g, 10mmol) and imidazole (2.04 g, 30 mmol) in DMSO (50 mL) was addedL-proline (0.345 g, 3 mmol), CuI (1.14 g, 6 mmol) and K₂CO₃ (2.76 g, 20mmol). The resultant mixture was stirred under N₂ at 100° C. overnight,cooled to room temperature, filtered, and concentrated in vacuum. Theresidue was dissolved in ethyl acetate and saturated aqueous NaHCO₃ wasadded till pH=8.5. The mixture was filtered and the resultant aqueouslayer was extracted with EA (5 times). The combined organic layers werewashed with brine, dried over MgSO₄, concentrated and purified by silicagel column chromatography (DCM:MeOH=30:1-20:1) to afford3-(5-(4-(1H-imidazol-1-yl)phenyl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoateas a pale solid (1.6 g, 36%).

Step 5: Synthesis of3-(5-(4-(1H-imidazol-1-yl)phenyl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid (5E, Ar1=4-carbamoyl-2-methylphenyl, R═H)

To a solution of compound3-(5-(4-(1H-imidazol-1-yl)phenyl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoate(22.0 g, 48.3 mmol) in THF/H₂O (v/v=1/1, 220 mL) was added LiOH.H₂O(4.15 g, 96.6 mmol). The reaction solution was stirred at roomtemperature for 5 h. The THF was removed under reduced pressure and theaqueous solution was acidified with 10% HCl to pH=5. The solid wasfiltered and recrystallized from THF and water [1:1 (v/v)] to afford3-(5-(4-(1H-imidazol-1-yl)phenyl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid as a yellow solid (11.35 g, 55%).

Representative Procedure for Scheme 6: Synthesis of3-(5-(benzo[d][1,3]dioxol-5-yl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid Step 1: Synthesis of 3-methyl-4-(1H-pyrrol-1-yl)benzamide (6A)

The 2,5-dimethoxy-tetrahydrofuran (106 g, 80 mmol) was added to thesolution of 4-amino-3-methylbenzamide (100 g, 66.7 mmol) in AcOH (300mL). The mixture was stirred at 80° C. for about 1.5 h and then cooledto room temperature. The solution of Na₂CO₃ was added dropwise at 0° C.and extracted with ethyl acetate for three times. The combined organiclayers were washed with brine, dried over Na₂SO₄, concentrated andwashed with petroleum ether. The resultant solid was filtrated and driedto afford 3-methyl-4-(1H-pyrrol-1-yl)benzamide as a pale solid (89.7 g,yield 67%).

Step 2: Synthesis of 4-(2-formyl-1H-pyrrol-1-yl)-3-methylbenzonitrile(6B)

POCl₃ (65 g, 427 mmol) was added to DMF (34 mL) at 0° C. for 30 min.After addition, the mixture was stirred at room temperature for 1.5 h,and then cooled to 0° C. A solution of3-methyl-4-(1H-pyrrol-1-yl)benzamide (6A) (42.7 g, 213.5 mmol) in DMF(150 mL) was added at 0° C. and the resultant mixture was stirred atroom temperature for 20 min, and then heated to 80° C. for 1 h. Thesolution was cooled to room temperature and then sat. Na₂CO₃ was addedat 0° C. until pH=8. The mixture was extracted with ethyl acetate threetimes. The combined organic layers were washed with sat. NaHCO₃ andbrine, dried over Na₂SO₄, concentrated and purified by silica gel columnchromatography (PE:EA=10:1) to afford4-(2-formyl-1H-pyrrol-1-yl)-3-methylbenzonitrile as a yellow solid (30.5g, yield 68%).

Step 3: Synthesis of ethyl3-(1-(4-cyano-2-methylphenyl)-1H-pyrrol-2-yl)acrylate (6C)

Method A

The mixture of 4-(2-formyl-1H-pyrrol-1-yl)-3-methylbenzonitrile (15 g,71.4 mmol) and (carbethoxymethylene)-triphenylphosphorane (27.5 g, 78.6mmol) in toluene was heated to 100° C. overnight. Then it was cooled toroom temperature, concentrated and purified by silica gel columnchromatography (PE:EA=5:1) to afford ethyl3-(1-(4-cyano-2-methylphenyl)-1H-pyrrol-2-yl)acrylate as a yellow oil(19.8 g, 98%).

Method B

To a mixture of 4-(2-formyl-1H-pyrrol-1-yl)-3-methylbenzonitrile (24.5g, 116.7 mmol), DMAP (2.9 g, 23.3 mmol) and potassium monoethyl malonate(99.2 g, 583.3 mmol) in DMF (600 mL) was added AcOH (35.0 g, 583.3 mmol)and piperidine (29.8 g, 350 mmol). The resultant mixture was heated to80° C. and stirred for 48 h. The reaction mixture was poured into cooledwater and extracted with ethyl acetate (800 mL×3). The combined organiclayers were washed with sat. NaHCO₃ and brine, dried over Na₂SO₄,concentrated and purified by silica gel column chromatography(PE:EA=5:1) to afford ethyl3-(1-(4-cyano-2-methylphenyl)-1H-pyrrol-2-yl)acrylate as a yellow oil(21.8 g, 67%).

Step 4: Synthesis of ethyl3-(1-(4-cyano-2-methylphenyl)-1H-pyrrol-2-yl)propanoate (6D)

To a solution of ethyl3-(1-(4-cyano-2-methylphenyl)-1H-pyrrol-2-yl)acrylate (6C) (8.0 g, 28.6mmol) in ethanol was added 10% Pd/C (0.8 g). The mixture was stirredunder 1 atm of H₂ for 30 min at room temperature and filtered. Theresultant filtrate was concentrated to dryness affording the crudeproduct of ethyl 3-(1-(4-cyano-2-methylphenyl)-1H-pyrrol-2-yl)propanoate(7.5 g), which was used for the next step without further purification:LC-MS m/z 283.0 [M+H]⁺, purity 68%.

Step 5: Synthesis of ethyl3-(5-bromo-1-(4-cyano-2-methylphenyl)-1H-pyrrol-2-yl)propanoate (6E)

NBS (4.76 g, 1 equiv) was added portionwise to a solution of ethyl3-(1-(4-cyano-2-methylphenyl)-1H-pyrrol-2-yl)propanoate in DMF at 0° C.during 45 min. After addition, the mixture was stirred at roomtemperature for 30 min, then poured into water, and extracted with ethylacetate for three times. The combined organic layers were washed withbrine, dried over Na₂SO₄, concentrated and purified by silica gel columnchromatography (PE:EA=15:1) to afford ethyl3-(5-bromo-1-(4-cyano-2-methylphenyl)-1H-pyrrol-2-yl)propanoate as awhite solid.

Step 6: Synthesis of ethyl3-(5-(benzo[d][1,3]dioxol-5-yl)-1-(4-cyano-2-methylphenyl)-1H-pyrrol-2-yl)propanoate

To a suspension of ethyl3-(5-bromo-1-(4-cyano-2-methylphenyl)-1H-pyrrol-2-yl)propanoate (400 mg,0.665 mmol), 3,4-methylenedioxylphenylboric acid (143 mg, 0.864 mmol),sodium bicarbonate (560 mg, 5.32 mmol) in solvent (4 mL) was addedPd(PPh₃)₄ (60 mg, 0.199 mmol). The reaction was degassed and heated toreflux for 5 h. TLC showed that the reaction was completed. Water (4 mL)was added and the mixture was extracted with ethyl acetate (5 mL×3). Thecombined organic layers were dried with magnesium sulfate, filtered andevaporated to obtain a brown oil, which was purified by silica gelcolumn chromatography to afford ethyl3-(5-(benzo[d][1,3]dioxol-5-yl)-1-(4-cyano-2-methylphenyl)-1H-pyrrol-2-yl)propanoateas a colorless oil (308 mg, 69%).

Step 7: and Step 8: Synthesis of3-(5-(benzo[d][1,3]dioxol-5-yl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid

To a mixture of ethyl3-(5-(benzo[d][1,3]dioxol-5-yl)-1-(4-cyano-2-methylphenyl)-1H-pyrrol-2-yl)propanoate(100 mg, 0.249 mmol) and potassium carbonate (52 mg, 0.373 mmol) in DMSO(1 mL) was added 30% aqueous H₂O₂ (28.2 mg, 0.249 mmol). The resultantmixture was stirred at room temperature for 2 h. TLC showed the reactionwas completed. Water (7 mL) was added and white solid precipitated. Thesuspension was centrifuged and the aqueous phase was discarded. Theresultant solid was dried in vacuum to afford the amide intermediate asa white solid (85 mg, yield 81%). To the mixture of this intermediate inH₂O (0.6 mL) and THF (0.6 mL) was added LiOH.H₂O (10 mg, 0.238 mmol).The reaction mixture was stirred at room temperature overnight. THF wasevaporated in vacuum. The residue was acidified to pH=4 with 5%hydrochloric acid, centrifuged and dried to afford3-(5-(benzo[d][1,3]dioxol-5-yl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid as a white solid (46 mg, overall yield 47%).

Scheme 7-Scheme 8 intentionally omitted.

Representative Procedure for Scheme 9a: Synthesis of3-[1[(4-carbamoyl-2-methyl-phenyl)-5-(4-pyrazole-1-yl)-1Hpyrrol-2-yl]-propionicacid Synthesis of3-[1-(4-carbamoyl-2-methyl-phenyl)-5-(4-pyrazole-1-yl-phenyl)-1H-pyrrol-2-yl]-propionicacid ethyl ester (9a-B, Ar=1H-pyrazol-1-yl)

N,N-dimethyl-cyclohexane-1,2-diamine (11 mg, 0.08 mmol) was dissolved inDMSO and degassed by bubbling argon through the solution for 2 minutes.The resulting solution was then added to a mixture of3-[1-(4-carbamoyl-2-methyl-phenyl)-5-(4-iodo-phenyl)-1H-pyrrole-2-yl]-propionicacid ethyl ester (which was prepared according to the first 3 steps ofScheme 1, R2=4-iodo-phenyl, and R1=4-carbamoyl-2-methylphenyl) (150 mg,0.29 mmol), pyrazole (500 mg, 7.5 mmol), copper iodide (11 mg, 0.06mmol), and potassium carbonate (86 mg (0.61 mmol) and the resultingreaction mixture again degassed for 2 minutes by bubbling argon gasthrough the solution. The reaction mixture was then submitted tomicrowave irradiation for 30 minutes at 120° C. The reaction mixture wasthen added to water (10 mL), extracted into ethyl acetate (3×10 mL). Theethyl acetate extracts were combined, washed with water (5 mL) and thenbrine (5 mL). The organic layer was then dried over MgSO₄.Chromatography (5 g silica sep-pak cartridge) with dichloromethane then1% methanol in dichloromethane yielded pure intermediate3-[1[(4-carbamoyl-2-methyl-phenyl)-5-(4-pyrazole-1-yl)-1Hpyrrol-2-yl]-propionicacid ethyl ester (26 mg, 20%).

Synthesis of3-(5-(4-(1H-pyrazol-1-yl)phenyl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid (9a-C, Ar=1H-pyrazol-1-yl)

3-[1-(4-carbamoyl-2-methyl-phenyl)-5-(4-pyrazole-1-yl-phenyl)-1H-pyrrol-2-yl]-propionicacid ethyl ester (24 mg, 0.06 mmol) was hydrolyzed using the proceduredescribed above in the final step of scheme 1 to give the titlecompound,3-[1[(4-carbamoyl-2-methyl-phenyl)-5-(4-pyrazole-1-yl)-1Hpyrrol-2-yl]-propionicacid (18 mg, 75%).

Representative Procedure for Scheme 9b: Synthesis of3-[1-(4-carbamoyl-2-methyl-phenyl)-5-(5-imidazole-1-yl-thiophene-2-yl)-1H-pyrrole-2-yl]-propionicacid Synthesis of3-[1-(4-carbamoyl-2-methyl-phenyl)-5-(5-imidazole-1-yl-thiophene-2-yl)-1H-pyrrole-2-yl]-propionicacid ethyl ester

Prepared using same protocol as Step 1 of Scheme 9a except starting withethyl3-(5-(5-bromothiophen-2-yl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoate(which was prepared according to the first 3 steps of Scheme 1,R₂=5-bromothiophen-2-yl, and R₁=4-carbamoyl-2-methylphenyl).

Synthesis of3-[1-(4-carbamoyl-2-methyl-phenyl)-5-(5-imidazole-1-yl-thiophene-2-yl)-1H-pyrrole-2-yl]-propionicacid

3-[1-(4-carbamoyl-2-methyl-phenyl)-5-(5-imidazole-1-yl-thiophene-2-yl)-1H-pyrrole-2-yl]-propionicacid ethyl ester was hydrolyzed according to the procedure described inthe final step of scheme 1 to give the title compound3-[1-(4-carbamoyl-2-methyl-phenyl)-5-(5-imidazole-1-yl-thiophene-2-yl)-1H-pyrrole-2-yl]-propionicacid.

Scheme 10-Scheme 18 intentionally omitted.

Representative Procedure for Scheme 19: Synthesis of3-[5-Benzothiazol-6-yl-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl]-propionicacid (19F, Ar2=benzothiazol-6-yl) Synthesis of Benzothiazole-6-carbonylchloride (19A, Ar2=benzothiazol-6-yl)

Under a nitrogen atmosphere, benzothiazole-6-carboxylic acid (1.014 g,5.6 mmol) was dissolved in methylene chloride (25 mL). Five drops ofN,N-dimethylforamide was added. Oxalyl chloride (0.5 mL, 5.6 mmol) wasslowly added. After 2 hrs, the reaction was heated to 30° C. for 16 hrs.The reaction was concentrated in vacuo to yield benzothiazole-6-carbonylchloride (1.665 g, quant., light yellow powder)

Synthesis of 7-(Benzothiazole-6-carbonyl)-1,4-dioxa-spiro[4.5]decan-8-one (19B, Ar2=benzothiazol-6-yl)

Under a nitrogen atmosphere, lithium hexamethyldisilazide (2.4 mL, 2.4mmol) was mixed with THF (5 mL). The reaction was cooled to −78° C.1,4-cyclohexane-dione monoethylene acetal (374 mg, 2.4 mmol), dissolvedin THF (2 mL) was slowly added via dropping funnel. The reaction wasstirred for 20 min at −78° C. It was then cannulated to a flask, cooledat −78° C., containing benzothiazole-6-carbonyl chloride (498 mg, 2.52mmol) dissolved in THF (5 mL). After the addition, the reaction wasstirred at −78° C. for 1 hr, and then allowed to warm to roomtemperature. After 12 h, water (30 mL) was added and extracted withethyl acetate (3×20 mL). The combined organic layers were washed with10% citric acid (20 mL), water (20 mL), bicarb (20 mL), and brine (20mL). It was then dried over Na₂SO₄, filtered and concentrated in vacuo.The crude material was purified by silica gel column (1:1 EtOAc/Hexanes)to yield 7-(Benzothiazole-6-carbonyl)-1, 4-dioxa-spiro[4.5]decan-8-one(271 mg, 35%, light yellow solid).

Synthesis of 3-[2-(3-Benzothiazol-6-yl-3-oxo-propyl)-[1,3]dioxolan-2-yl]-propionic acid ethyl ester (19C, Ar2=benzothiazol-6-yl)

Under a nitrogen atmosphere, 7-(Benzothiazole-6-carbonyl)-1,4-dioxa-spiro[4.5]decan-8-one (271 mg, 0.85 mmol) was dissolved inethanol (1 mL). 2.43 M sodium ethoxide solution (0.01 mL, 0.03 mmol) wasadded. After 12 hrs, reaction was concentrated in vacuo. The residue wasdiluted with 10 mL EtOAc/5 mL 10% citric acid. The layers wereseparated. The aqueous layer was further extracted with EtOAc (3×3 mL).The combined organic layers were washed with water (5 mL) and brine (5mL), dried over Na₂SO₄, filtered and concentrated in vacuo. The crudematerial was purified by silica gel column (40% EtOAc/hexanes) to yield3-[2-(3-Benzothiazol-6-yl-3-oxo-propyl)-[1, 3]dioxolan-2-yl]-propionicacid ethyl ester (100 mg, 38%, light yellow oil).

Synthesis of 7-Benzothiaol-6-yl-4,7-dioxo-heptanoic acid ethyl ester(19D, Ar2=benzothiazol-6-yl)

Under a nitrogen atmosphere,3-[2-(3-Benzothiazol-6-yl-3-oxo-propyl)-[1,3]dioxolan-2-yl]-propionicacid ethyl ester (19C) (100 mg, 0.28 mmol) was dissolved in THF (1 mL).3N HCl was added and stirred at room temperature. After 12 hrs, thereaction was diluted with water and extracted with EtOAc (3 times). Thecombined organic layers were washed with brine, dried over Na₂SO₄,filtered and concentrated in vacuo to give7-Benzothiaol-6-yl-4,7-dioxo-heptanoic acid ethyl ester (52 mg, 58%,dark red solid; ⅔ as ethyl ester, ⅓ as carboxylic acid).

Synthesis of3-[5-Benzothiazol-6-yl-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl]-propionicacid ethyl ester (19E, Ar2=benzothiazol-6-yl)

In a 4 mL vial, purged with nitrogen,7-Benzothiaol-6-yl-4,7-dioxo-heptanoic acid ethyl ester (52 mg, 0.16mmol) was dissolved in 2 mL ethanol. P-toluenesulfonic acid (pTSA) (9.9mg, 0.05 mmol) and 4-amino-3-methyl benzamide (37 mg, 0.24 mmol) wereadded. The vial was capped tightly and heated to 80° C. in an oil bath.After the 12 hrs, the reaction was cooled and concentrated in vacuo. Thecrude material was dissolved in N,N-dimethylforamide (1 mL). Potassiumcarbonate (44 mg, 0.32 mmol) was added. Then iodoethane (0.01 mL, 0.17mmol) was added. The reaction was stirred at room temperature for 12hrs. The reaction was diluted with water and extracted with ethylacetate. The combined organic layers were washed with water, brine anddried over Na₂SO₄, filtered and concentrated in vacuo. The crude productwas purified by silica gel column (5% IPA/CH₂Cl₂) to give3-[5-Benzothiazol-6-yl-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl]-propionicacid ethyl ester (19E, Ar2=benzothiazol-6-yl) (42 mg, 73% over 2 steps,red solid).

Synthesis of3-[5-Benzothiazol-6-yl-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl]-propionicacid (19F, Ar2=benzothiazol-6-yl)

3-[5-Benzothiazol-6-yl-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl]-propionicacid ethyl ester (19E) (42 mg, 0.10 mmol) was hydrolyzed according tothe procedure described above in the final step of scheme 4, to give thetitle compound3-[5-Benzothiazol-6-yl-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl]-propionicacid (23 mg, 59%, light tan powder).

Representative Procedure for Scheme 20: Synthesis of3-(5-(4-(1H-imidazol-1-yl)phenyl)-1-(2-methyl-4-(methylsulfonamido)phenyl)-1H-pyrrol-2-yl)propanoicacid (20C, Ar2=4-(1H-imidazol-1-yl)phenyl) Synthesis of3-(5-(4-(1H-imidazol-1-yl)phenyl)-1-(4-amino-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid (20B, Ar2=4-(1H-imidazol-1-yl)phenyl)

3-(5-(4-(1H-imidazol-1-yl)phenyl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid (20A, prepared according to Scheme 5,Ar2=4-carbamoyl-2-methylphenyl) (3.88 g, 9.37 mmol) was added to aq.NaOH (4.12 g, 103.09 mmol, dissolving in 50 mL). Then 11% aq. NaClO(28.83 g, 42.17 mmol) was added dropwise. The resulting mixture was keptat 0-10° C. for 1 h, at 35° C. for 1 h and at 75° C. for 30 min. Aftercooling to room temperature, the reaction was acidified with 10%hydrochloric acid to pH=7.0 and filtered to remove the solid impurity.The filtrate was further acidified with 10% hydrochloric acid to pH=5.0and a new precipitate appeared. The precipitate was filtrated and driedto afford 20B, Ar2=4-(1H-imidazol-1-yl)phenyl as a gray powder (3.20 g,88%).

Synthesis of3-(5-(4-(1H-imidazol-1-yl)phenyl)-1-(2-methyl-4-(methylsulfonamido)phenyl)-1H-pyrrol-2-yl)propanoicacid (20C, Ar2=4-(1H-imidazol-1-yl)phenyl)

To a solution of pyridine (2 mL) and CH₃SO₂Cl/DCM (v/v=1/100, 5 mL) wasadded a solution of3-(5-(4-(1H-imidazol-1-yl)phenyl)-1-(4-amino-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid (20B) (250 mg, 0.74 mmol) in pyridine (2 mL) at 0° C. The mixturewas stirred at room temperature for 1 h. The solvents were removed underreduced pressure and the resulting solid was acidified with 10%hydrochloric acid to pH=5.0. The resulting precipitate was isolated bycentrifuge, rinsed with water, dried under reduced pressure to afford20C, Ar2=4-(1H-imidazol-1-yl)phenyl as a brown powder (40 mg, 13%).

Scheme 21-Scheme 32 intentionally omitted.

Representative Procedure for Scheme 33: Synthesis of3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-chloro-2-methoxyphenyl)-1H-pyrrol-2-yl)propanoicacid (33C, R1=4-carbamoyl-2-methylphenyl, R2=4-chloro, R3=methylSynthesis of ethyl3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-chloro-2-hydroxyphenyl)-1H-pyrrol-2-yl)propanoate(33A, R1=4-carbamoyl-2-methylphenyl, R2=4-chloro)

Prepared following scheme 1 thru 1C, R1=4-carbamoyl-2-methylphenyl,R2=4-chloro-2-hydroxyphenyl.

Synthesis of ethyl3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-chloro-2-methoxyphenyl)-1H-pyrrol-2-yl)propanoate(33B, R1=4-carbamoyl-2-methylphenyl, R2=4-chloro, R3=methyl)

Ethyl3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-chloro-2-hydroxyphenyl)-1H-pyrrol-2-yl)propanoate(300 mg, 0.704 mmol) was dissolved in acetone. Potassium carbonate (146mg, 1.056 mmol) and methyl iodide (299 mg, 2.112 mmol) was added andstirred at room temperature overnight. When TLC indicated that thereaction was complete, the mixture was filtered, evaporated in vacuo.The residue was partitioned between ethyl acetate (20 mL) and water (5mL). The organic phase was dried with magnesium sulfate, filtered andconcentrated to afford 33B, R1=4-carbamoyl-2-methylphenyl, R2=4-chloro,R3=methyl as a yellow oil (295 mg, yield 95%).

Synthesis of3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-chloro-2-methoxyphenyl)-1H-pyrrol-2-yl)propanoicacid (33C, R1=4-carbamoyl-2-methylphenyl, R2=4-chloro, R3=methyl)

Hydrolysis completed following final step of Scheme 5 to give the titlecompound.

Representative Procedure for Scheme 34 Synthesis of3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-(2-cyclopropyl-1H-imidazol-1-yl)phenyl)-1H-pyrrol-2-yl)propanoicacid (34C, Ar1-X=4-bromophenyl, Ar2 is 2-cylopropyl-1H-imidazol-1-yl,R1=4-carbamoyl-2-methylphenyl) Synthesis of ethyl3-(5-(4-bromophenyl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoate(34A, R1=4-carbamoyl-2-methylphenyl, Ar1-X=4-bromophenyl)

Prepared by Scheme 1, steps 1-3.

Synthesis of ethyl3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-(2-cyclopropyl-1H-imidazol-1-yl)phenyl)-1H-pyrrol-2-yl)propanoate(34B, Ar1-X=4-bromophenyl, Ar2 is 2-cyclopropyl-1H-imidazol-1-yl,R1=4-carbamoyl-2-methylphenyl

To a mixture of 34A (Ar2=4-bromophenyl) (455 mg, 1.0 mmol) and2-cyclopropyl-1H-imidazole (see Method 14 for synthesis) (324 mg, 3.0mmol, 3.0 eq) in NMP (4 mL) was added 8-hydroxyquinoline (22 mg, 0.15mmol, 0.15 eq), Cu₂O (282 mg, 0.1 mmol) and K₂CO₃ (166 mg, 1.2 mmol) andPEG-2000 (50 mg). The resultant mixture under N₂ was irradiated undermicrowave at 128° C. for 6.0 h, cooled to room temperature and dilutedwith THF (10 mL) and water (10 mL). The mixture was filtered and theresultant aqueous layer was extracted with EA (30 mL×5). The combinedorganic layers were washed with brine (20 mL), dried over MgSO₄,filtered, concentrated and purified by silica gel column chromatography(MeOH:CH₂Cl₂=1:15) to afford the desired compound as a yellow solid (190mg, yield 39%).

Synthesis of3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-(2-cyclopropyl-1H-imidazol-1-yl)phenyl)-1H-pyrrol-2-yl)propanoicacid (34C, Ar1-X=4-bromophenyl, Ar2 is 2-cyclopropyl-1H-imidazol-1-yl,R1=4-carbamoyl-2-methylphenyl)

Hydrolysis completed following final step of Scheme 5 to give the titlecompound.

Scheme 35 intentionally omitted.

Representative Procedure for Scheme 36:3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-(2-oxooxazolidin-3-yl)phenyl)-1H-pyrrol-2-yl)propanoicacid Synthesis of ethyl4,7-dioxo-7-(4-(2-oxooxazolidin-3-yl)phenyl)heptanoate

To a mixture of ethyl 7-(4-bromophenyl)-4,7-dioxoheptanoate ((36A, whereAr1-Br=4-bromophenyl, also see compound 5B, Scheme 5) (1.50 g, 4.4 mmol)and oxazolidin-2-one (575 mg, 6.6 mmol) in dioxane (5 mL) were addedL-proline (50 mg, 0.44 mmol), CuI (42 mg, 0.22 mmol) and K₂CO₃ (1.22 g,8.8 mmol). The resultant mixture was stirred under N₂ at 110° C. for 48h and then evaporated. The residue was diluted with EA/water (40 mL/40mL). The mixture was filtered and the resultant aqueous layer wasextracted with EA (30 mL×5). The combined organic layers were washedwith brine, dried over NaSO₄, concentrated and purified by silica gelcolumn chromatography (pure DCM to DCM:MeOH=30:1) to afford titledcompound as a white solid (158 mg, yield 10%).

Synthesis of ethyl3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-(2-oxooxazolidin-3-yl)phenyl)-1H-pyrrol-2-yl)propanoate

To a solution of ethyl4,7-dioxo-7-(4-(2-oxooxazolidin-3-yl)phenyl)heptanoate (158 mg, 0.43mmol) and 4-amino-3-methylbenzamide (130 mg, 0.68 mmol) in EtOH (1 mL)was added Zn(OTf)₂ (313 mg, 0.86 mmoL). The mixture was heated to 120°C. under microwave for 2 h. After evaporation under reduced pressure,the crude product was purified by silica gel column chromatography(DCM:MeOH=20:1) to afford the titled compound as a yellow solid (77 mg,yield 39%).

Synthesis of3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-(2-oxooxazolidin-3-yl)phenyl)-1H-pyrrol-2-yl)propanoicacid

To a solution of ethyl3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-(2-oxooxazolidin-3-yl)phenyl)-1H-pyrrol-2-yl)propanoate(67 mg, 0.15 mmol) in THF/H₂O (1 mL, v/v=1/1) was added lithiumhydroxide monohydrate (7 mg, 0.15 mmol). The mixture was stirred at roomtemperature for 6 h. THF was evaporated in vacuo. The residue wasacidified to pH=5 with 5% hydrochloric acid, concentrated and purifiedby prep-TLC to afford the titled compound as a brown solid (24 mg, yield39%).

Representative Procedure for Scheme 36A: Synthesis of ethyl7-(3-fluoro-4-(1H-imidazol-1-yl)phenyl)-4,7-dioxoheptanoate (R═H)

Ethyl 7-(3,4-difluorophenyl)-4,7-dioxoheptanoate (351 mg) was treatedwith imidazole (241 mg) and pyridine (395 mg) in DMSO (3 mL) at 150° C.over 7 h with a micro-wave heating. The resultant mixture was dilutedwith water (12 mL) and was extracted with EtOAc (20 mL×3). After removalthe solvents, the mixture was purified by flash silica gelchromatography, eluting with EtOAc, to afford the desired product -ethyl7-(3-fluoro-4-(1H-imidazol-1-yl)phenyl)-4,7-dioxoheptanoate (279 mg,68%) as light brown solids.

Scheme 37-Scheme 38 intentionally omitted.

Synthesis of 39A

To a mixture of 16-4 (Method 16) (200 mg, 0.419 mmol), NaBH₃CN (53 mg,0.838 mmol), 37% HCHO (1.5 mL, 2.095 mmol) in CH₃CN (5 mL) was addedAcOH (0.5 mL). After stirred at room temperature overnight, the solutionwas concentrated and diluted with water (15 mL), extracted with ethylacetate (10 mL×4). The organic phase was separated and dried, purifiedwith prep-TLC (PE:EA=1:1) to afford 39A as a yellow oil (97 mg, 49%).

Synthesis of 39B

Followed the procedure described in the last two steps of Scheme 6(steps 7 and 8), with a purification of the final product by prep-HPLC.

Synthesis of ethyl3-(5-(4-chloro-2-formamidophenyl)-1-(4-cyano-2-methylphenyl)-1H-pyrrol-2-yl)propanoate(40A)

A mixture of Ac₂O (301 mg, 2.948 mmol) and HCO₂H (226 mg, 4.914 mmol)was stirred at 55° C. for 5 min. The mixture was added to the solutionof 16-4 (see method 16 for synthesis) (300 mg, 0.737 mmol) in THF (6 mL)and stirred at 55° C. for 10 min. TLC showed the reaction was complete.The volatiles were removed under reduced pressure, and the residue wasdissolved in EA (50 mL), washed with sat. NaHCO₃ (10 mL×3) and brine (10mL). The organic layer was dried over anhydrous Na₂SO₄, filtered andconcentrated to afford the crude product as a yellow solid (320 mg,yield: 99%) which used for the next step directly.

Synthesis of3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-chloro-2-formamidophenyl)-1H-pyrrol-2-yl)propanoicacid (40B)

See methodology described in the last steps of Scheme 6 (6F→6H).

The Following Methods were Used to Prepare Intermediates that were Usedin Schemes Above as Noted in the Table.

Method 1-Method 11 intentionally omitted.

Compound 12-2

A solution of 12-1 (12.3 g, 0.06 mmol), Bis(pinacolato)diboron (18.3 g,0.072 mol), KOAc (11.75 g, 0.12 mmol) and Pd(dppf)₂Cl₂.DCM (2.0 g, 2.45mmol) in dioxane/H₂O (v/v=9/1, 100 mL) was stirred at 80° C. overnight.TLC showed that the reaction was complete. The mixture was evaporated toafford a brown oil. Water (60 mL) was added and the mixture wasextracted with ethyl acetate (60 mL×3). The combined organic layers weredried over MgSO₄, filtered, concentrated and purified by silica gelcolumn chromatography (PE:EA=10:1) to afford 12-2 as a yellow solid (9.1g, 60%).

Method 13-Method 14 intentionally omitted.

Compound 15-1

To a stirred suspension of 3-bromophenol (50 g, 0.29 mol) in pyridine(200 mL) and dichloromethane (100 mL) was added dropwise acetyl chloride(25 mL, 0.35 mol) at 0° C. and the mixture was stirred 18 h at roomtemperature. LC-MS showed that the reaction was complete. Pyridine anddichloromethane was evaporated in vacuo. Water (600 mL) was added andacidified with hydrochloric acid at pH 2. The reaction mixture wasextracted with ethyl acetate (500 mL×3) and the organic phase was driedover anhydrous sodium sulfate, filtrated, concentrated and purified bycolumn chromatography (PE:EA=60:1) to afford compound 15-1 as acolorless liquid (46 g, 74%).

Compound 15-2

To a stirred suspensions of compound 15-1 (46 g, 0.0.21 mol) andanhydrous aluminum chloride power (57 g, 0.42 mol) was heated to 160° C.for 3 h. The mixture reaction was cooled to room temperature and ice(200 g) and water (800 mL) was poured and purified with hydrochloricacid at pH 7. the reaction was extracted with ethyl acetate (500 mL×3)and the organic phase was washed with saturated sodium bicarbonate,dried over anhydrous sodium sulfate, filtrated, concentrated andpurified by column chromatography (PE:EA=60:1) to afford compound 15-2as a light green solid (35.1 g, 76%).

Compound 15-3

To a suspension of compound 15-2 (25 g, 0.12 mol) and potassiumcarbonate (24 g, 0.18 mol) in anhydrous DMF (20 mL) was added to MeI(22.6 mL, 0.23 mol) and the mixture reaction was stirred at roomtemperature overnight. LCMS showed that the reaction was complete. Thenwater (300 mL) was poured and the mixture was extracted with ethylacetate and the organic phase was (200 mL×3) and the organic phase waswashed saturated sodium chloride, dried over anhydrous sodium sulfate,filtrated, concentrated to afford compound 15-3 as a colorless solid(26.1 g, 98%).

Compound 16-2

To a solution of 16-1 (6.50 g, 27.66 mmol) and NiCl₂ (7.80 g, 55.3 mmol)in EtOH (50 mL) was added NaBH₄ (5.60 g, 138.3 mmol) slowly. Theresultant mixture was stirred at 0° C. for 2 h, filtered andconcentrated under reduced pressure. The residue was dissolved withethyl acetate (200 mL), washed with water (50 mL×3), dried over Na₂SO₄,concentrated and purified by silica gel column (PE:EA=5:1) to afford16-2 as a dark solid (3.778 g, yield 67%).

Compound 16-3

A solution of 16-2 (3.778 g, 18.43 mmol), Bis(pinacolato)diboron (8.5 g,33.17 mol), KOAc (3.2 g, 36.86 mmol) and Pd(dppf)₂Cl₂.DCM (500 mg, 0.92mmol) in DMSO (50 mL) was stirred at 85° C. for 2.5 h. TLC showed thatthe reaction was complete. Water (60 mL) was added and the mixture wasextracted with ethyl acetate (60 mL×3). The combined organic layers weredried over Na₂SO₄, filtered, concentrated and purified by silica gelcolumn (PE:EA=10:1) to afford 16-3 as a yellow solid (5.0 g, yield100%).

Compound 16-4

To a solution of 16-3 (7.0 g, 27.7 mmol), Na₂CO₃ (11.75 g, 110.8 mmol)and 6E (ethyl3-(5-bromo-1-(4-cyano-2-methylphenyl)-1H-pyrrol-2-yl)propanoate, seeScheme 6) (10 g, 21.4 mmol) in DMSO (30 mL) was added Pd(PPh₃)₄ (3.0 g,8.31 mmol). After having been degassed and recharged with nitrogen, thereaction mixture was stirred at 80° C. overnight. TLC showed thereaction was complete. After cooling to room temperature, water (50 mL)was added and extracted with ethyl acetate (50 mL×4). The combinedorganic layers were dried over Na₂SO₄, filtered, concentrated andpurified by silica gel column (PE:EA=3:1) to afford 16-4 as a yellowsolid (3.10 g, yield 27%).

Method 17 intentionally omitted.

Compound 18-2

Prepared following the same procedure described in Method 12, withdioxane as the solvent and purified by column chromatography (PE:EA=5:1)to give 35% yield of desired.

Compound 19-2

To a solution of 3-acetylthiophene (2.52 g, 20 mmol, 1.0 eq) in HOAc (50mL) was added NaOAc (2.46 g, 30 mmol, 1.5 eq) followed by bromine (3.2g, 20 mmol, 1.0 eq) dropwise over 30 min. The mixture was allowed tostir at room temperature overnight. Water (150 mL) was added and thereaction mixture was stirred for 2 h. The resulting solid was collectedby filtration, rinsed with water (10 mL) and PE (20 mL) and dried toafford 19-2 as a brown solid (1.52 g, yield 37%).

Method 20-Method 22 intentionally omitted.

Representative Example for Method 23: Synthesis ofN-(4-aminophenyl)methanesulfonamide (23-3, R═H)

Compound 23-2, R═H

To a solution of pyridine (50 mL) and MsCl (15.86 g, 139.13 mmol) in DCM(150 mL) was added the solution of 4-nitrobenzenamine (16.0 g, 115.94mmol) in pyridine (100 mL) at 0° C. The mixture was stirred at roomtemperature for 4 h. The volatiles were removed under reduced pressure.The residue was rinsed with water (200 mL×3) and dried under reducedpressure to afford the title compound as a yellow powder (23.20 g, yield95%).

Compound 23-3, R═H

To a solution of 23-2, R═H (23.0 g, 106.48 mmol) in MeOH (100 mL) wasadded 10% Pd/C (3.0 g) purged with N₂. Then a solution of HCO₂NH₄ (67.0g, 1.06 mol) in MeOH (500 mL) was added gradually under ice-water bathduring 5 min. After addition, the mixture was warmed to 45° C. andstirred overnight and filtered. The filtrate was evaporated underreduced pressure to afford yellow solid which was washed with EA (500mL×3). The combined organic layers were evaporated under reducedpressure, purified by silica gel column chromatography (PE:EA=1:2) toafford N-(4-aminophenyl)methanesulfonamide as a yellow solid (9.80 g,yield 49%).

Compound 24-1

To a solution of 3-chlorothiophene (4.80 g, 40.48 mmol) in THF (50 mL)was added BuLi (2.5N in hexane, 17.9 mL) at −30° C. After addition, themixture was stirred for 30 min at −10° C., and then cooled to −45° C.N-methoxy-N-methyl acetamide (55.0 g, 48.8 mmol) was added and allowedto warm to room temperature during 40 min and maintained for anadditional 20 min. Brine (80 mL) was added to quench the reaction,extracted with EA (60 mL×3). The combined organic layers were dried overanhydrous Na₂SO₄, filtered, concentrated to afford 24-1 (˜80% pure) as ayellow oil (6.80 g) which used for the next step directly.

Compound 25-1

To a suspension of N,O-dimethylhydroamine hydrochloride (100 g, 1026mmol) in DCM (1000 mL) was added triethylamine (300 mL, 2052 mmol) at 0°C. Acetyl chloride was added dropwise to the suspension for 2 h at 0° C.When the addition was complete, the mixture was allowed to warm to roomtemperature and stirred for 2 h. The mixture was washed with brine (1L), 1 N HCl (500 mL), brine (200 mL) respectively and dried withmagnesium sulfate, filtered and concentrated to afford brown oil, whichwas purified by distillation to afford 25-1 as a colorless liquid (65 g,61%).

Compound 25-2

To a solution of thiophene (84 g, 1.0 mol) in chloroform (34 mL) wasadded dropwise bromine at room temperature for 3 h. When the additionwas complete, the mixture was stirred at room temperature overnight. Themixture was heated to 50° C. for 3 h. The reaction mixture was washedwith 1M NaOH (aq. 100 mL), brine (100 mL×2) respectively. The organicphase was dried with anhydrous sodium sulfate, filtered and concentratedto afford light yellow oil, which was solidified in methanol (100 mL).The solid was filtered and dried in vacuo to afford 25-2 (89 g, 56%).

Compound 25-3

25-2 (9.5 g, 30 mmol) was dissolved in anhydrous THF (100 mL) and cooledto −78° C. To the above solution was added dropwise n-BuLi (8 mL, 21mmol) for 30 min and stirred for 30 min. 25-1 was added dropwise at −78°C., stirred for 30 min and allowed to warm to room temperature beforequenching with saturated ammonium chloride. The organic phase wasseparated and washed with brine, dried with anhydrous Na₂SO₄, filteredand concentrated to afford yellow oil, which was purified by columnchromatography (elution: PE/EA=10/1) afford 25-3 as a yellow solid (2.3g, 28%).

Compound 25-4

To a solution of 25-3 (2.4 g, 8.5 mmol) in methanol (35 mL) was addedtrimethyl orthofomate (15 mL) and TsOH (300 mg, 1.7 mmol). The solutionwas heated to reflux for 10 h. Methanol was evaporated in vacuo and theresidue was partitioned between EA (300 mL) and 5% sodium bicarbonate(100 mL). The organic phase was separated, dried with anhydrous sodiumsulfate, filtered and concentrated to afford 25-4 as a yellow oil, whichwas used directly for next step (2.3 g, 82%).

Compound 25-5

To a solution of 25-4 (6.0 g, 18.3 mmol) in DMF (75 mL) was added sodiummethoxide (9.9 g, 183 mmol), cuprous oxide (1.5 g, 18.3 mmol) and sodiumiodide (2.8 g, 18.3 mmol). The mixture was heated to 100° C. for 4 h.TLC indicated that the reaction was complete and the reaction wasquenched with brine (250 mL). The solid was filtered and the filtratewas extracted with ethyl acetate (100 mL×3). The combined organic layerswere dried with anhydrous sodium sulfate, filtered and concentrated toafford brown oil, which was purified by column chromatography (elution:PE/EA=3/1) to afford 25-5 as a light yellow oil (1.2 g, 23%).

Compound 25-6

To a solution of 25-5 (1.2 g, 4.29 mmol) in DCM (8 mL) and water (10 mL)was added trifluoroacetic acid (10 mL). The reaction mixture was stirredat room temperature for 4 h. Saturated sodium bicarbonate (10 mL) wasadded and the organic phase was separated, dried with anhydrous sodiumsulfate, filtered and concentrated to afford brown oil, which waspurified by column chromatography (elution: PE/EA=10/1) to afford 25-6as a light yellow solid (750 g, 74%).

Method 26 intentionally omitted.

Compound 27-2

Ac₂O (16 ml, 0.16 mol) was added to the solution of 27-1 (20 g, 0.107mol) in 80 ml of pyridine. The mixture was stirred at room temperaturefor 2 hours. Then EtOH (40 ml) was added and the solid was isolated byfiltration and washed with EtOH to give 27-2 as a brown solid (10.3 g,yield 56%).

Compound 27-3

Compound 27-2 (10 g, 43.6 mmol) was added to a flask containing 1 N NaOH(36 ml) and the mixture was stirred at room temperature for 3 hours. Thesolvent was removed and the residue was washed with EtOH. 27-3 wasisolated by filtration as a pale solid (8.8 g, yield 88%).

Compound 27-4

Compound 27-3 (16 g, 63.7 mmol) and DMF (20 ml) were added to a flaskand then SOCl₂ (18.4 g, 155 mol) was added dropwise at −30-40° C. Whenthe addition was complete, the mixture was stirred at room temperaturefor 2 hours. Then the mixture was added to ice slowly and solidappeared. The solid was isolated by filtration and dried to give 27-4 asa pale solid (6.0 g, yield 38%).

Compound 27-5

The solution of 27-4 (6.0 g, 24.2 mmol) in 50 ml of THF was added to 50ml of NH₄OH at 0° C. dropwise. The mixture was stirred at roomtemperature for 1 h. The solvent was removed under reduced pressure andthe residue was extracted with EA (30 ml×4). The organic layer was driedover Na₂SO₄ and filtered, concentrated to give 27-5 as a pale solid (5.1g, yield 93%).

Compound 27-6

A mixture of 27-5 (5.1 g, 22.3 mmol), HCl (2 N, 76.5 ml) and EtOH (100ml) was refluxed overnight. Then the mixture was neutralized withNa₂CO₃(aq) to PH=8. The mixture was extracted with EA (80 ml×4), driedover Na₂SO₄, and concentrated to give 27-6 as a pale solid (4.9 g, yield100%).

Compound 28-1

To a solution of 3-chlorothiophene (6.52 g, 55 mmol) in CHCl₃ (30 mL)and AcOH (30 mL) was added NBS (9.80 g, 55 mmol). The mixture was heatedat reflux for 1.5 h, then cooled to room temperature. Water (70 mL) wasadded and the mixture was extracted with CHCl₃ (30 mL×2). The combinedorganic layers were washed with sat. NaHCO₃ (40 mL) and brine (30 mL),dried over anhydrous Na₂SO₄, filtered, concentrated to afford 28-1 as abrown oil (10.02 g, quantitative yield) which used for the next stepdirectly.

Synthesis of 1-(5-bromo-4-chlorothiophen-2-yl)ethanone (28-2)

To a mixture of 28-1 (10.0 g, 50.6 mmol) and AlCl₃ (8.09 g, 60.7 mmol)in DCM (120 mL) was added dropwise acetyl chloride (4.76 g, 60.7 mmol)during 5 min at 0° C. After addition, the mixture was stirred overnightat room temperature, washed with diluted hydrochloride acid (1.2N, 150mL) and brine (150 mL), dried over anhydrous Na₂SO₄, filtered,concentrated and purified by silica gel column chromatography(PE/EA=20/1 to 3/1) to afford 28-2 as a brown solid (8.0 mg, yield:66%).

Synthesis of 1-(4-chlorothiophen-2-yl)ethanone (28-3)

To a solution of 28-2 (3.20 mg, 13.36 mmol) in EtOH (70 mL) was added10% Pd/C (2.50 g) and AcONa (1.10 g, 13.36 mmol). The reaction mixturewas stirred under a hydrogen atmosphere at room temperature for 3 h,filtered, and the filtrate was concentrated. The resultant residue wasdissolved in EA (100 mL) washed with sat. NaHCO₃ (40 mL) and brine (30mL), dried over anhydrous Na₂SO₄, filtered, concentrated and purified bysilica gel column chromatography (PE/EA=30/1 to 5/1) to afford 28-3 as ayellow oil (1.32 g, yield: 62%).

Method 29-Method 32 intentionally omitted.

Synthesis of Compound 33-2

A mixture of HCO₂H (644 mg, 14 mmol) and Ac₂O (1.16 g, 11.4 mmol) washeated to 55° C. for 2 h and then cooled to room temperature. THF (1 mL)and 33-1 (880 mg, 4.38 mmol) in THF (1 mL) was added stepwise and theresultant mixture was continually stirred at room temperature for 3 h.After evaporation, the residue was extracted with EA (5 mL×3). Theorganic phase was successively washed with sat. aqueous sodiumbicarbonate (10 mL) and sodium chloride (10 mL), dried over anhydroussodium sulfate, filtrated and concentrated to afford 33-2 as a liquid(845 mg, yield: 85%), which was used directly for the next step.

Synthesis of Compound 33-3

To a mixture of 33-2 (845 mg, 4.38 mmol), KOAc (726 mg, 7.4 mmol),B(pin)₂ (1.41 g, 5.6 mmol) and dioxane was added Pd(dppf)₂Cl₂ (20 mg,0.02 mmol). After having been degassed and recharged with nitrogen, themixture was refluxed at 90° C. overnight. TLC showed that the reactionwas complete. Water (10 mL) was added and the mixture was extracted withethyl acetate (10 mL×3). The combined organic layers were dried overNa₂SO₄, filtered, concentrated and purified by silica gel columnchromatography (PE:EA=4:1) to afford 33-3 as a colorless solid (220 mg,yield: 29%).

Method 34 intentionally omitted.

Method 35: Synthesis of 4-amino-benzenesulfonamide

Followed procedure/scheme described in Method 27, Synthesis of4-amino-3-methylbenzenesulfonamide.

Method 36-40 intentionally omitted.

Compound 41-2

To a solution of 41-1 (2.0 g, 13.32 mmol) in acetone (25 mL) was addedNBS (2.37 g, 13.32 mmol) and 1M HCl aq. (0.13 mL, 0.13 mmol). Thereaction mixture was stirred at room temperature for 3 h, and thenconcentrated to dryness under reduced pressure. The residue wasdissolved with PE (40 mL) the resultant precipitate was filtered anddried in vacuum to afford 41-2 as a white solid (2.90 g, yield: 95%).

Example 2: GSNOR Assays

Various compounds were tested in vitro for their ability to inhibitGSNOR activity. Representative compounds and their corresponding GSNORactivity are described in a paragraph before Table 1 above. GSNORexpression and purification is described in Biochemistry 2000, 39,10720-10729.

GSNOR Fermentation:

Pre-cultures were grown from stabs of a GSNOR glycerol stock in 2XYTmedia containing 100 ug/ml ampicillin after an overnight incubation at37° C. Cells were then added to fresh 2XYT (4 L) containing ampicillinand grown to an OD (A₆₀₀) of 0.6-0.9 at 37° C. before induction. GSNORexpression was induced with 0.1% arabinose in an overnight incubation at20° C.

GSNOR Purification:

E. coli cell paste was lysed by nitrogen cavitation and the clarifiedlysate purified by Ni affinity chromatography on an AKTA FPLC (AmershamPharmacia). The column was eluted in 20 mM Tris pH 8.0/250 mM NaCl witha 0-500 mM imidazole gradient. Eluted GSNOR fractions containing theSmt-GSNOR fusion were digested overnight with Ulp-1 at 4° C. to removethe affinity tag then re-run on the Ni column under the same conditions.GSNOR was recovered in the flowthrough fraction and for crystallographyis further purified by Q-Sepharose and Heparin flowthroughchromatography in 20 mM Tris pH 8.0, 1 mM DTT, 10 uM ZnSO₄.

GSNOR Assay: Procedure

GSNO and Enzyme/NADH Solutions are made up fresh each day. The Solutionsare filtered and allowed to warm to room temperature. GSNO Solution: 100mM NaPO4 (pH 7.4), 0.480 mM GSNO. 396 μL of GSNO Solution is added to acuvette followed by 8 μL of test compound in DMSO (or DMSO only for fullreaction control) and mixed with the pipette tip. Compounds to be testedare made up at a stock concentration of 10 mM in 100% DMSO. 2 foldserial dilutions are done in 100% DMSO. 8 μL of each dilution are addedto an assay so that the final concentration of DMSO in the assay is 1%.The concentrations of compounds tested range from 100 to 0.003 μM.Enzyme/NADH Solution: 100 mM NaPO4 (pH 7.4), 0.600 mM NADH, 1.0 μg/mLGSNO Reductase. 396 μL of the Enzyme/NADH Solution is added to thecuvette to start the reaction. The cuvette is placed in the Cary 3EUV/Visible Spectrophotometer and the change in 340 nm absorbance/min at25° C. is recorded for 3 minutes. The assays are done in triplicate foreach compound concentration. IC50's for each compound are calculatedusing the standard curve analysis in the Enzyme Kinetics Module ofSigmaPlot.

Final assay conditions: 100 mM NaPO4, pH 7.4, 0.240 mM GSNO, 0.300 mMNADH, 0.5 μg/mL GSNO Reductase and 1% DMSO. Final volume: 800μL/cuvette.

Example 3: GSNOR Inhibition Assay in an In Vivo Animal Model

To demonstrate the influence of GSNOR inhibition, a mouse model ofasthma was used that was similar to a model previously shown to beinfluenced by GSNO reductase and bioavailable SNO's (Que et al.,Science, 2005). Que et al. demonstrated that following ova-albumin (OVA)challenge, wild type mice exhibiting bronchial reactivity have increasedlevels of GSNOR and have lungs that were depleted of SNO's. In contrastto wild-type mice, Que et al. demonstrated that mice with a geneticdeletion of GSNOR increased lung SNO's and were protected from OVAinduced airway hyper-reactivity.

In an effort to determine if similar observations would manifest ifGSNOR were inhibited pharmacologically by a GSNOR inhibitor, an OVAmouse model (i.e., the wild-type model of Que et al.) was used. In thisstudy, OVA sensitized mice were administered 1 mg/kg, 10 mg/kg or 30mg/kg of Compound 1 intravenously at 24 hours prior to being placed in awhole body plethysmograph (Buxco Research Systems, Wilmington, N.C.) andprovided with fresh air.

Subject animals were then challenged with an aerosol of increasingdosages of the bronchoconstrictive agent methacholine, a pharmacologicagent commonly used in determining the degree of bronchialhyper-reactivity in experimental subjects. In this study mice wereexposed to an increasing concentration of methacholine, each dose beingpresented for 3 minutes, during which time readings were taken. Doses ofmethacholine were 0 mg/ml, 5 mg/ml, 20 mg/ml, and 50 mg/ml. The degreeof bronchial hyper-reactivity was measured as the ‘Enhanced Pause’(Penh), a unit-less index of airway hyper-reactivity (Dohi et al., LabInvest. 79(12):1559-1571, 1999).

The administration of Compound 1 produced lower broncho-constrictiveresponses in these test animals compared with vehicle-only dosedanimals. These results are consistent with a greater level of bioactiveSNO's available to counter the bronchoconstrictive methacholinechallenge.

Example 4: Efficacy of GSNORi in Experimental Asthma

Experimental Asthma Model:

A mouse model of ovalbumin (OVA)-induced asthma was used to screen GSNORinhibitors for efficacy against methacholine (MCh)-inducedbronchoconstriction/airway hyper-reactivity. This is a widely used andwell characterized model that presents with an acute, allergic asthmaphenotype with similarities to human asthma. Efficacy of GSNORinhibitors were assessed using a prophylactic protocol in which GSNORinhibitors were administered prior to challenge with MCh.Bronchoconstriction in response to challenge with increasing doses ofMCh was assessed using whole body plethysmography h, (P_(enh); Buxco).The amount of eosinophil infiltrate into the bronchoaveolar lavage fluid(BALF) was also determined as a measure of lung inflammation. The effectof GSNOR inhibitors were compared to vehicles and to Combivent (inhaled;IH) as the positive control.

Materials and Methods

Allergen Sensitization and Challenge Protocol

OVA (500 μg/ml) in PBS was mixed with equal volumes of 10% (w/v)aluminum potassium sulfate in distilled water and incubated for 60 min.at room temperature after adjustment to pH 6.5 using 10 N NaOH. Aftercentrifugation at 750×g for 5 min, the OVA/alum pellet was resuspendedto the original volume in distilled water. Mice received anintraperitoneal (IP) injection of 100 μg OVA (0.2 mL of 500 μg/mL innormal saline) complexed with alum on day 0. Mice were anesthetized byIP injection of a 0.2-mL mixture of ketamine and xylazine (0.44 and 6.3mg/mL, respectively) in normal saline and were placed on a board in thesupine position. Two hundred fifty micrograms (100 μl of a 2.5 mg/ml) ofOVA (on day 8) and 125 μg (50 μl of 2.5 mg/ml) OVA (on days 15, 18, and21) were placed on the back of the tongue of each animal.

Pulmonary Function Testing (Penh)

In vivo airway responsiveness to methacholine was measured 24 h afterthe last OVA challenge in conscious, freely moving, spontaneouslybreathing mice with whole body plethysmography using a Buxco chamber(Wilmington, N.C.). Mice were challenged with aerosolized saline orincreasing doses of methacholine (5, 20 and 50 mg/mL) generated by anultrasonic nebulizer for 2 min. The degree of bronchoconstriction wasexpressed as enhanced pause (P_(enh)), a calculated dimensionless value,which correlates with the measurement of airway resistance, impedance,and intrapleural pressure in the same mouse. P_(enh) readings were takenand averaged for 4 min. after each nebulization challenge. P_(enh) wascalculated as follows: P_(enh)=[(T_(e)/T_(r)−1)×(PEF/PIF)], where T_(e)is expiration time, T_(r) is relaxation time, PEF is peak expiratoryflow, and PIF is peak inspiratory flow×0.67 coefficient. The time forthe box pressure to change from a maximum to a user-defined percentageof the maximum represents the relaxation time. The T_(r) measurementbegins at the maximum box pressure and ends at 40%.

Eosinophil Infiltrate in BALF

After measurement of airway hyper-reactivity, the mice wereexsanguinated by cardiac puncture, and then BALF was collected fromeither both lungs or from the right lung after tying off the left lungat the mainstem bronchus. Total BALF cells were counted from a 0.05 mLaliquot, and the remaining fluid is centrifuged at 200×g for 10 min at4° C. Cell pellets were resuspended in saline containing 10% BSA withsmears made on glass slides. Eosinophils were stained for 5 min. with0.05% aqueous eosin and 5% acetone in distilled water, rinsed withdistilled water, and counterstained with 0.07% methylene blue.

GSNOR Inhibitors and Controls

GSNOR inhibitors were reconstituted in phosphate buffered saline (PBS),pH 7.4, at concentrations ranging from 0.00005 to 3 mg/mL. GSNORinhibitors were administered to mice (10 mL/kg) as a single dose eitherintravenously (IV) or orally via gavage. Dosing was performed from 30min. to 24 h prior to MCh challenge. Effect of GSNOR inhibitors werecompared to PBS vehicle dosed in the same manner.

Combivent was used as the positive control in all studies. Combivent(Boehringer Ingelheim) was administered to the lung using the inhalerdevice supplied with the product, but adapted for administration tomice, using a pipet tip. Combivent was administered 48 h, 24 h, and 1 hprior to MCh challenge. Each puff (or dose) of Combivent provided a doseof 18 μg ipatropium bromide (IpBr) and 103 μg albuterol sulfate orapproximately 0.9 mg/kg IpBr and 5 mg/kg albuterol.

Statistical Analyses

Area under the curve values for P_(enh) across baseline, saline, andincreasing doses of MCh challenge were calculated using GraphPad Prism5.0 (San Diego, Calif.) and expressed as a percent of the respective (IVor orally administered) vehicle control. Statistical differences amongtreatment groups and the respective vehicle control group within eachstudy were calculated using one-way ANOVA, Dunnetts (JMP 8.0, SASInstitute, Cary, N.C.). A p value of <0.05 among the treatment groupsand the respective vehicle control group was considered significantlydifferent.

Results:

Compound 1 Results

Compound 1 administered intravenously (IV) was efficacious againstexperimental asthma as noted by attenuation of methacholine (MCh)induced bronchoconstriction and pulmonary inflammation. Significantefficacy with Compound 1 was observed with a single IV dose of 0.01mg/kg at 24 h prior to MCh. The area under the curve (AUC) for Penhresponse reported as percent of vehicle control (AUC=100%) was 42.1±2.8%(p<0.0001). Eosinophil infiltration into the bronchoaveolar lavage fluid(BALF) was reduced by 98% (p<0.0001). Significant efficacy with Compound1 was also observed as early as 1 h (AUC=76.4±6.6; p=0.0082) and up to48 h (AUC=64.4±55; p=<0.0001) prior to MCh at a single IV dose of 0.1mg/kg. The ED50, the dose of Compound 1 demonstrating 50% reduction inPenh response, was 0.011±0.003 mg/kg.

Compound 2 Results

Compound 2 administered intravenously (IV) was efficacious againstexperimental asthma as noted by attenuation of methacholine (MCh)induced bronchoconstriction. Significant efficacy with Compound 2 wasobserved with a single IV dose of 0.01, 0.1, and 1 mg/kg at 24 h priorto MCh. The area under the curve (AUC) for Penh response reported aspercent of vehicle control (AUC=100%) was 65.3±6.5% (p=0.0002);50.5±6.3% (p<0.0001); and 41.7±5.2% (p<0.0001) for 0.01, 0.1, and 1mg/kg, respectively, of Compound 2.

Compound 3 Results

Compound 3 administered intravenously (IV) was efficacious againstexperimental asthma as noted by attenuation of methacholine (MCh)induced bronchoconstriction and pulmonary inflammation. Significantefficacy with Compound 3 was observed with a single IV dose of 1 mg/kgat 24 h prior to MCh. The area under the curve (AUC) for Penh responsereported as percent of vehicle control (AUC=100%) was 71.0±8.6%(p=0.0051). Eosinophil infiltration into the bronchoaveolar lavage fluid(BALF) was reduced by 46% (p=0.0002).

Compound 6 Results

Compound 6 administered intravenously (IV) or orally was efficaciousagainst experimental asthma as noted by attenuation of methacholine(MCh) induced bronchoconstriction and pulmonary inflammation.Significant efficacy with Compound 6 was observed with a single IV doseof 1 mg/kg at 24 h prior to MCh. The area under the curve (AUC) for Penhresponse reported as percent of vehicle control (AUC=100%) was 65.3±5.9%(p=0.0001). Eosinophil infiltration into the bronchoaveolar lavage fluid(BALF) was reduced by 92% (p<0.0001). Significant efficacy with Compound6 was also observed with a single oral dose of 30 mg/kg at 24 h prior toMCh. The area under the curve (AUC) for Penh response reported aspercent of vehicle control (AUC=100%) was 24.6±3.0% (p<0.0001).Eosinophil infiltration into the bronchoaveolar lavage fluid (BALF) wasreduced by 100% (p=0.0004).

Compound 7 Results

Compound 7 administered intravenously (IV) was efficacious againstexperimental asthma as noted by attenuation of methacholine (MCh)induced bronchoconstriction. Significant efficacy with Compound 7 wasobserved with a single IV dose of 0.1 and 1 mg/kg at 24 h prior to MCh.The area under the curve (AUC) for Penh response reported as percent ofvehicle control (AUC=100%) was 56.1±2.2% (p<0.0001) and 50.4±3.7%(p<0.0001) 0.1 and 1 mg/kg of Compound 7, respectively.

Compound 26 Results

Compound 26 administered intravenously (IV) or orally was efficaciousagainst experimental asthma as noted by attenuation of methacholine(MCh) induced bronchoconstriction and pulmonary inflammation.Significant efficacy with Compound 26 was observed with a single IV doseof 0.1, 1, and 10 mg/kg at 24 h prior to MCh. The area under the curve(AUC) for Penh response reported as percent of vehicle control(AUC=100%) was 64.2±7.6% (p=0.0007); 60.2±7.9% (p=0.0002); and 40.7±2.4%(p<0.0001) for 0.1 mg/kg, 1 mg/kg, and 10 mg/kg, respectively, ofCompound 26. Eosinophil infiltration into the bronchoaveolar lavagefluid (BALF) was reduced by 79% (p=0.0064); 100% (p=0.0007); and 100%(p=0.0007) for 0.1 mg/kg, 1 mg/kg, and 10 mg/kg, respectively, ofCompound 26. Significant efficacy with Compound 26 was also observed asearly as 30 min. (AUC=35.2±9.3; p<0.0001) prior to MCh at a single IVdose of 10 mg/kg. Eosinophil infiltration into the BALF was reduced by94% (p<0.0001). Significant efficacy with Compound 26 was also observedwith a single oral dose of 30 mg/kg at 24 h prior to MCh. The area underthe curve (AUC) for Penh response reported as percent of vehicle control(AUC=100%) was 26.7±1.4% (p<0.0001). Eosinophil infiltration into thebronchoaveolar lavage fluid (BALF) was reduced by 100% (p=0.0019).

Compound 33 Results

Compound 33 administered intravenously was efficacious againstexperimental asthma as noted by attenuation of methacholine (MCh)induced bronchoconstriction and pulmonary inflammation. Significantefficacy with Compound 33 was observed with a single IV dose of 1 mg/kgat 24 h prior to MCh. The area under the curve (AUC) for Penh responsereported as percent of vehicle control (AUC=100%) was 72.9±8.7%(p=0.0089). Eosinophil infiltration into the bronchoaveolar lavage fluid(BALF) was reduced by 61% (p<0.0001).

Compound 67 Results

Compound 67 administered intravenously (IV) was efficacious againstexperimental asthma as noted by attenuation of methacholine (MCh)induced bronchoconstriction and pulmonary inflammation. Significantefficacy with Compound 67 was observed with a single IV dose of 1 mg/kgat 24 h prior to MCh. The area under the curve (AUC) for Penh responsereported as percent of vehicle control (AUC=100%) was 78.7±8.1%(p=0.0323). Eosinophil infiltration into the bronchoaveolar lavage fluid(BALF) was reduced by 63% (p<0.0001).

It will be apparent to those skilled in the art that variousmodifications and variations can be made in the methods and compositionsof the present invention without departing from the spirit or scope ofthe invention.

The invention claimed is:
 1. A method of treatment of a disorder whichcomprises administering to a patient in need thereof, a therapeuticallyeffective amount of a compound of Formula (I)

wherein: Ar is selected from the group consisting of phenyl andthiophen-yl; R₁ is selected from the group consisting of unsubstitutedimidazolyl, substituted imidazolyl, chloro, bromo, fluoro, hydroxy, andmethoxy; R₂ is selected from the group consisting of hydrogen, methyl,chloro, fluoro, hydroxy, methoxy, ethoxy, propoxy, carbamoyl,dimethylamino, amino, formamido, and trifluoromethyl; and X is selectedfrom the group consisting of CO and SO₂; or a pharmaceuticallyacceptable salt thereof, and wherein the compound is administered with asecondary active agent.
 2. The method of claim 1 wherein the secondaryactive agent is selected from the group consisting of an NO donor, otherNO bioactivity generating compounds, an NO releaser, a chemotherapeuticagent, an agent that imposes nitrosative or oxidative stress, aphosphodiesterase inhibitor, and a β-agonist.
 3. The method of claim 1wherein R₁ is selected from the group consisting of unsubstitutedimidazolyl and substituted imidazolyl.
 4. The method of claim 3 whereinthe substituted imidazolyl group is substituted with C₁-C₆ alkyl.
 5. Themethod of claim 3 wherein ArR₁R₂ is selected from the group consistingof:

wherein R₃ is selected from H, methyl, and ethyl.
 6. The method of claim3 wherein the compound of formula I is selected from the groupconsisting of3-(5-(4-(1H-imidazol-1-yl)phenyl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(5-(1H-imidazol-1-yl)thiophen-2-yl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-(2-methyl-1H-imidazol-1-yl)phenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-(4-methyl-1H-imidazol-1-yl)phenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-(2-ethyl-1H-imidazol-1-yl)phenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(4-(1H-imidazol-1-yl)thiophen-2-yl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(5-(2-methyl-1H-imidazol-1-yl)thiophen-2-yl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(3-fluoro-4-(1H-imidazol-1-yl)phenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(3-fluoro-4-(2-methyl-1H-imidazol-1-yl)phenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-(2-methyl-1H-imidazol-1-yl)thiophen-2-yl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(2-methoxy-4-(2-methyl-1H-imidazol-1-yl)phenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(4-(1H-imidazol-1-yl)-2-methoxyphenyl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(5-(2-methyl-1H-imidazol-1-yl)thiophen-3-yl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(5-(2-ethyl-1H-imidazol-1-yl)thiophen-2-yl)-1H-pyrrol-2-yl)propanoicacid; and3-(5-(5-(2-methyl-1H-imidazol-1-yl)thiophen-2-yl)-1-(2-methyl-4-sulfamoylphenyl)-1H-pyrrol-2-yl)propanoicacid.
 7. The method of claim 1 wherein ArR₁ are selected from the groupconsisting of: 4-chlorophenyl, 3-chlorophenyl, 4-bromophenyl,3-bromophenyl, 4-fluorophenyl, 3-fluorophenyl, 4-hydroxyphenyl,4-methoxyphenyl, 3-methoxyphenyl, 2-methoxyphenyl,4-chlorothiophen-2-yl, 5-chlorothiophen-2-yl, 3-bromothiophen-2-yl,4-bromothiophen-2-yl, 5-bromothiopheny-2-yl, and 5-bromothiophen-3-yl.8. The method of claim 1 wherein the compound of formula I is selectedfrom the group consisting of3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-hydroxyphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(5-bromothiophen-2-yl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-methoxyphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(4-bromophenyl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(3-chloro-4-methoxyphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(3-fluoro-4-methoxyphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(3-chloro-4-hydroxyphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-methoxy-3-methylphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(3-methoxyphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(4-amino-3-chlorophenyl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(3,4-difluorophenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(2,4-difluorophenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-chlorophenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(4-bromothiophen-2-yl)-1H-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-fluoro-3-methoxyphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-carbamoyl-3-fluorophenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-methoxy-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-chloro-2-fluorophenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-fluorophenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-fluoro-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-chloro-2-methoxyphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(2-chloro-4-methoxyphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(2-ethoxy-4-fluorophenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-methoxy-2-(trifluoromethyl)phenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-fluoro-2-methoxyphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-chloro-3-fluorophenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-chloro-2-ethoxyphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(5-bromo-2-methoxyphenyl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(4-bromo-2-methoxyphenyl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-chloro-2-hydroxyphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(5-bromothiophen-3-yl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-hydroxy-3-methylphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(2-carbamoyl-4-chlorophenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(2-methoxyphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(2,4-dimethoxyphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-chloro-2-propoxyphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-hydroxy-2-methoxyphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-chloro-2-(dimethylamino)phenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(5-chlorothiophen-2-yl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-chloro-2-formamidophenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(3-chlorothiophen-2-yl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-formamido-2-methoxyphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(3-bromo-5-methoxythiophen-2-yl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(1-(4-carbamoyl-2-methylphenyl)-5-(4-chlorothiophen-2-yl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(5-bromo-4-chlorothiophen-2-yl)-1-(4-carbamoyl-2-methylphenyl)-1H-pyrrol-2-yl)propanoicacid; and3-(5-(4-bromothiophen-2-yl)-1-(2-methyl-4-sulfamoylphenyl)-1H-pyrrol-2-yl)propanoicacid.
 9. The method of claim 1 wherein the disorder is selected from thegroup consisting of pulmonary disorders and chronic inflammatorydiseases.
 10. The method of claim 9 wherein the pulmonary disorder isselected from the group consisting of asthma, chronic obstructivepulmonary disease (COPD), and cystic fibrosis.
 11. The method of claim10 wherein the pulmonary disorder is cystic fibrosis.
 12. The method ofclaim 9 wherein the chronic inflammatory disease is selected from thegroup consisting of inflammatory bowel disease (IBD), Crohn's disease,colitis, psoriasis, and AIDS related dimentia.
 13. A method of treatmentof a disorder which comprises administering to a patient in needthereof, a therapeutically effective amount of a compound of Formula (I)

wherein: Ar is selected from the group consisting of phenyl andthiophen-yl; R₄ is selected from the group consisting of unsubstitutedimidazolyl and substituted imidazolyl; R₅ is selected from the groupconsisting of hydrogen, fluoro, hydroxy, and methoxy; R₆ is selectedfrom the group consisting of hydrogen, chloro, bromo, and fluoro; R₇ isselected from the group consisting of hydrogen, and methyl; and R₈ isselected from the group consisting of CONH₂, SO₂NH₂, and NHSO₂CH₃; or apharmaceutically acceptable salt thereof, and wherein the compound isadministered with a secondary active agent.
 14. The method of claim 13wherein the secondary active agent is selected from the group consistingof an NO donor, other NO bioactivity generating compounds, an NOreleaser, a chemotherapeutic agent, an agent that imposes nitrosative oroxidative stress, a phosphodiesterase inhibitor, and a β-agonist. 15.The method of claim 13 wherein the substituted imidazolyl group issubstituted with C₁-C₆ alkyl.
 16. The method of claim 13 wherein ArR₄R₅is selected from the group consisting of:

wherein R₉ is selected from H, methyl, and ethyl.
 17. The method ofclaim 13 wherein the compound of formula II is selected from the groupconsisting of3-(5-(5-(2-methyl-1H-imidazol-1-yl)thiophen-2-yl)-1-(4-sulfamoylphenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(5-(2-methyl-1H-imidazol-1-yl)thiophen-2-yl)-1-(2-methyl-4-(methylsulfonamido)phenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(4-(1H-imidazol-1-yl)phenyl)-1-(2-methyl-4-(methylsulfonamido)phenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(4-(2-methyl-1H-imidazol-1-yl)phenyl)-1-(2-methyl-4-(methylsulfonamido)phenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(4-(2-methyl-1H-imidazol-1-yl)thiophen-2-yl)-1-(2-methyl-4-(methylsulfonamido)phenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(5-(2-methyl-1H-imidazol-1-yl)thiophen-2-yl)-1-(2-methyl-4-(methylsulfonamido)phenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(4-(2-methyl-1H-imidazol-1-yl)thiophen-2-yl)-1-(4-(methylsulfonamido)phenyl)-1H-pyrrol-2-yl)propanoicacid;3-(5-(2-methoxy-4-(2-methyl-1H-imidazol-1-yl)phenyl)-1-(4-(methylsulfonamido)phenyl)-1H-pyrrol-2-yl)propanoicacid; and3-(5-(4-(2-methyl-1H-imidazol-1-yl)phenyl)-1-(4-(methylsulfonamido)phenyl)-1H-pyrrol-2-yl)propanoicacid.
 18. The method of claim 13 wherein the disorder is selected fromthe group consisting of pulmonary disorders and chronic inflammatorydiseases.
 19. The method of claim 18 wherein the pulmonary disorder isselected from the group consisting of asthma, chronic obstructivepulmonary disease (COPD), and cystic fibrosis.
 20. The method of claim19 wherein the pulmonary disorder is cystic fibrosis.
 21. The method ofclaim 18 wherein the chronic inflammatory disorder is selected from thegroup consisting of inflammatory bowel disease (IBD), Crohn's disease,colitis, psoriasis, and AIDS related dimentia.